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Transcriptional expression of survivin and its splice variants in brain tumors in humans
Abstract
Survivin, one of the apoptosis inhibitor proteins, has been detected in most cancers in humans. In addition, two splice variants (survivin-2B and survivin-deltaEx3) have been identified. The authors investigated the transcription levels of survivin messenger (m)RNA and its splice variants in nine tumor cell lines, including gliomas, and in 25 brain tumor samples, by performing quantitative reverse transcription-polymerase chain reaction. The correlation between transcript expression levels and pathological findings were also analyzed.
Transcription levels were measured using primer pairs specific for survivin and either of its splice variants and were normalized to the glyceraldehyde 6-phosphate dehydrogenase. Among the tumor cell lines tested, glioblastoma cell lines showed the highest levels of survivin expression. Among brain tumor samples studied, survivin was preferentially expressed in malignant brain tumors and gliomas. The relative expression level of survivin-deltaEx3/survivin was significantly higher in malignant than in benign brain tumor samples. Expression patterns were dominant for survivin-deltaEx3 in malignant brain tumors and dominant for survivin-2B in benign ones. A significant linear correlation between survivin mRNA expression and MIB-1 labeling index was demonstrated in all brain tumor samples.
The authors' results indicate that quantifying the levels of survivin and its splice variants is useful for the prediction of the cell biological malignancy of gliomas, independent of their pathological features.
... Moreover, overexpression of survivin is significantly associated with tumorigenesis and progression of gliomas, as well as the poor prognosis of patients with gliomas. [27][28][29][30][31][32][33] Nevertheless, the relation of survivin expression with proliferation, apoptosis, and angiogenesis of gliomas is still unclear. ...
... 39 Several reports have shown that mRNA and protein expression levels of survivin are markedly increased in gliomas by RT-PCR, Western blot, and immunohistochemistry; moreover, survivin overexpression is also correlated with pathologic grade and the poor prognosis of this malignancy. [27][28][29][30][31][32][33] In our previous study, survivin mRNA expression was detected by RT-PCR and similar results were obtained (data not shown herein). ...
... The results were consistent with previous research. [27][28][29][30][31][32][33] It is noteworthy that we found survivin protein located in the cytoplasm of glioma cells, as well as in the nucleus of glioma cells. This finding was not completely in accord with past reports. ...
Background:
An unbalance of cell proliferation and cell apoptosis is an important mechanism in carcinogenesis, and angiogenesis also plays a crucial role in tumorigenesis. Recently, survivin has been identified as an important member of the inhibitor of apoptosis protein (IAP) family. Although it has been shown that survivin is highly expressed in gliomas, and is associated with tumorigenesis, progression, and poor prognosis of gliomas, as yet the relation of survivin expression with proliferation, apoptosis, and angiogenesis of gliomas it is still unclear.
Methods:
Eighty-three cases of brain glioma were chosen and protein expressions of survivin and proliferating cell nuclear antigen (PCNA) in glioma cells and Factor VIII-related antigen (FVIII-RAg) in vascular endothelial cells were investigated by immunohistochemistry. Apoptotic cells of brain glioma were screened by TdT-mediated dUTP nick end-labeling (TUNEL), and survivin immunoreactivity score (IRS), proliferative index (PI), apoptotic index (AI), overall daily growth (ODG), and microvessel density (MVD) in brain gliomas were measured.
Results:
The survivin IRS, PI, AI, ODG, and MVD of brain gliomas were 3.75 +/- 3.89, 28.39 +/- 19.49%, 1.00 +/- 0.80%, 12.19 +/- 10.21%, and 62.75 +/- 31.50, respectively, and all of them increased markedly with an increase in the pathologic grade of brain gliomas (P < 0.001 for all). PI, ODG, and MVD in the survivin-positive group were significantly higher than those in the survivin-negative group (P < 0.001 for all). PI, ODG, and MVD were positively correlated with survivin IRS (P < 0.001 for all). Although there was no significant difference between AI in the survivin-positive group or in the survivin-negative group (P = 0.108), AI was inversely correlated with survivin IRS (P = 0.005).
Conclusions:
Survivin is overexpressed in brain gliomas, which may play an important role in malignant proliferation, antiapoptosis, and angiogenesis of brain gliomas.
... Human and non-human primates have splice variants which have additional sequences (for example, exon 2B or exon 3B) 29 , while, murine survivin variants are mainly deletion splicing isoforms (for example, survivin140 (WT), survivin121 and survivin40), and survivin-2B expression has never been reported in mice 31 . Survivin-2B has already been reported in many types of human malignant tumors: soft tissue sarcoma, gastric carcinoma, brain tumor, hepatocellular carcinoma, non-small-cell lung cancer (NSCLC) and acute myeloid leukemia (AML) 30,[32][33][34][35][36][37] . In the present study, we were able to detect survivin-2B expression in RA patients' sera and synovial tissues. ...
... The function of survivin-2B has been controversial. The expression levels of survivin-2B in solid tumors were often negatively correlated with the clinical stages of malignant tumors and the recurrence rates 33,34,36 . Therefore, these authors postulated that survivin-2B might diminish survivin-WT function in a dominant negative manner and could have pro-functions in solid tumors. ...
Survivin is an independent prognostic factor for joint destruction in rheumatoid arthritis (RA). However, the expression and function of survivin in RA synoviocytes remain unclear. We certified the expression of survivin in RA synovial tissues and performed the experiment using RA fibroblast-like synoviocytes (RA-FLS) treated with siRNA. As a result, the expression levels of wild type (WT) survivin and the 2B splice variants in RA synovial tissues were higher than those in osteoarthritis tissue samples, and, these variants were highly expressed in RA-FLS. The expression levels of survivin-WT and -2B in the RA-FLS were upregulated by PDGF. Treatment with siRNA against survivin-2B led to decreased viability of PDGF-treated RA-FLS due to cell cycle suppression and apoptosis promotion, while the siRNA against all survivin isoforms did not affect the viability. Moreover, an overexpression of survivin-2B in RA-FLS led to cell proliferation through cell cycle activation and by conferring resistance to apoptosis. In conclusion, survivin-2B has an important role in RA-FLS proliferation. These data suggest that survivin-2B might contribute to rheumatoid synovial hyperplasia, and have the potential as a novel therapeutic target for RA.
... Reports describing the clinico-pathologic relevance of survivin 2B expression in cancer are conflicting ( Table 2). While some authors report a stage dependent upregulation of 2B expression in colorectal carcinoma [3] as well as a grade dependent 2B upregulation in diffusely infiltrating astrocytoma [2], associated with adverse survival outcomes in both instances, others have linked high levels of 2B expression with positive clinical outcomes in the same cancers [40,41]. Specifically, increased levels of 2B expression have been associated with benign, rather than malignant brain tumors [40] and a significant correlation to a better prognosis in colorectal carcinoma [41]. ...
... While some authors report a stage dependent upregulation of 2B expression in colorectal carcinoma [3] as well as a grade dependent 2B upregulation in diffusely infiltrating astrocytoma [2], associated with adverse survival outcomes in both instances, others have linked high levels of 2B expression with positive clinical outcomes in the same cancers [40,41]. Specifically, increased levels of 2B expression have been associated with benign, rather than malignant brain tumors [40] and a significant correlation to a better prognosis in colorectal carcinoma [41]. In fact, about half of the studies investigating expression of 2B in cancer, report an inverse correlation between expression levels of this splice variant and adverse clinical outcomes (Table 2). ...
... Here, we analyzed the gene expression of Survivin and its splice variants: Survivin-ΔEx3 and Survivin-2B, as well as of the Cdk1 gene in GBM tumor samples, GBM cell lines, and normal samples by using quantitative qRT-PCR. To date, only one successful quantitative analysis of Survivin splice variants using TaqMan qRT-PCR in GBM samples has been reported [37]. Like Yamada et al. [37], we foundFig. ...
... To date, only one successful quantitative analysis of Survivin splice variants using TaqMan qRT-PCR in GBM samples has been reported [37]. Like Yamada et al. [37], we foundFig. 6 M4N causes apoptotic cell death, diminishes mitotic index (MI), and alters the cell cycle of glioblastoma cell lines. ...
Glioblastoma (GBM), one of the most malignant human neoplasias, responds poorly to current treatment modalities, with temozolomide (TMZ) being the drug most frequently used for its treatment. Tetra-O-methyl Nordihydroguaiaretic Acid (M4N) is a global transcriptional repressor of genes dependent on the Sp1 transcription factor, such as Survivin and Cdk1. In the present study we evaluated the gene expression of Survivin, its spliced variants and Cdk1 in GBM samples and cell lines. Moreover, we investigated the effects of M4N combined or not with TMZ and/or radiation on GBM primary cultures and cell lines. qRT-PCR assays were performed to determine the Survivin-spliced variants and Cdk1 gene mRNA expression in GBM tumor samples and cell lines. Cell proliferation was measured by XTT assay and cell cycle and apoptosis were determined by flow cytometry. Drug combination analyses using different schedules of administration (simultaneous and sequential) were performed on GBM cell lines and primary cultures based on the Chou-Talalay method. For clonogenic survival, doses of 2, 4, and 6 Gy of gamma radiation. were used. All Survivin-spliced variants and the Cdk1 gene were expressed in GBM samples (n = 16) and cell lines (n = 6), except the Survivin-2B variant that was only expressed in GBM cell lines. M4N treatment down regulated the expression of Cdk1, Survivin and the Survivin-ΔEx3 variant, while the Survivin-2B variant was up-regulated. M4N decreased the cell proliferation separately and synergistically with TMZ, and enhanced the effects of radiation, mainly when associated with TMZ. M4N also induced apoptotic cell death, decreased the mitotic index and arrested the cell cycle mainly in the G2/M phase. Our results suggest a potential clinical application of M4N in combination with TMZ and radiation for GB treatment.
... The mRNA expression of survivin 2B has been assessed in different cancers (14,(46)(47)(48). However contrasting results were obtained regarding the correlation between survivin 2B expression levels and clinicopathological markers (14,(46)(47)(48)(49). ...
... The mRNA expression of survivin 2B has been assessed in different cancers (14,(46)(47)(48). However contrasting results were obtained regarding the correlation between survivin 2B expression levels and clinicopathological markers (14,(46)(47)(48)(49). These contrasting results are mainly due to the use of non-specific primers to amplify specifically survivin 2B, or the use of semi-quantitative analysis of gel images obtained by RT-PCR (14,49). ...
To study the role of survivin and its splice variants in taxane-resistant ovarian cancer.
We assessed the mRNA levels of survivin splice variants in ovarian cancer cell lines and ovarian tumor samples. siRNAs targeting survivin were designed to silence all survivin splice variants (T-siRNA) or survivin 2B (2B-siRNA) in vitro and orthotopic murine models of ovarian cancer. The mechanism of cell death was studied in taxane-resistant ovarian cancer cells and in tumor sections obtained from different mouse tumors.
Taxane-resistant ovarian cancer cells express higher survivin mRNA levels than their taxane-sensitive counterparts. Survivin 2B expression was significantly higher in taxane-resistant compared with -sensitive cells. Silencing survivin 2B induced growth inhibitory effects similar to silencing total survivin in vitro. In addition, survivin 2B-siRNA incorporated into DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) nanoliposomes resulted in significant reduction in tumor growth (P < 0.05) in orthotopic murine models of ovarian cancer, and these effects were similar to T-siRNA-DOPC. The antitumor effects were further enhanced in combination with docetaxel chemotherapy (P < 0.01). Finally, we found a significant association between survivin 2B expression and progression-free survival in 117 epithelial ovarian cancers obtained at primary debulking surgery.
These data identify survivin 2B as an important target in ovarian cancer and provide a translational path forward for developing new therapies against this target.
... Therapie darstellen. Mit EGFRvIII [123], gp240 [124], Tenascin [125], Survivin [126], SART1 [127], IL-13Rα2 [128], TRP1 und 2, gp100 sowie MAGE-1 und MAGE-3 [129] wurden bereits Kandiaten-TAA identifiziert. Die ersten Ergebnisse von klinischen ...
... Das TAA und die Antigenbeladung unreifer DC sind also essentiell für die Induktion einer Gliomzell-spezifischen T-Zell-Antwort. Obwohl für Gliome zahlreiche TAA beschrieben wurden (EGFRvIII [123, 175], gp240 [124], AIM-2 [176], GLEA2 [177], Tenascin [125], Survivin [126, 178], SART1 [127], IL-13Rα2 [128], TRP1 und 2, gp100 sowie MAGE-1 und MAGE-3 [129, 179]), wurden in den bisherigen klinischen ...
Mit einem medianen Überleben von 14,6 Monaten ist die Prognose für das maligne Gliom noch immer schlecht und neue Therapieverfahren werden dringend benötigt. Die Vakzinierung mit Dendritischen Zellen (DC) stellt einen vielversprechenden Ansatz dar. DC sind spezialisierte antigenpräsentierende Zellen, die eine zentrale Rolle in der Initiationsphase der Immunantwort spielen. Voraussetzungen für die Vakzinierungstherapie ist eine ausreichende, nicht vom Tumor oder Vortherapie eingeschränkte Funktionalität des Immunsystems der Patienten und die in vitro Generierung funktionell kompetenter Dendritischer Zellen. Deshalb wurde im ersten Teil der Arbeit ein Immunstatus der Patienten erhoben. Gliompatienten wiesen einige Auffälligkeiten in der zellulären Zusammensetzung des Blutes auf. Besonders ausgeprägt war eine T-Lymphopenie, wobei T-Zell-Subpopulationen gleichermaßen betroffen waren. Jedoch zeigten die T-Zellen eine normale Antwort auf eine Lektinstimulation. Für die Vakzinierungstherapie werden Dendritische Zellen in vitro generiert, mit Tumorantigenen beladen und den Patienten verabreicht, mit der Vorstellung, dass sie im Patienten zur Entwicklung einer gegen den Tumor gerichteten Immunantwort führen. Mit einem Standardprotokoll zur in vitro Generierung Dendritischer Zellen lag der Anteil an CD83+, reifen Zellen, welche bei der Vakzinierung eingesetzt werden, nur bei 34,1%. Deshalb wurde ein serumfreies Zellkulturprotokoll etabliert, so dass die Anteil an reifen DC (64,3%) signifikant erhöht wurde, was sich funktionell in einem höheren allostimulatorischen Potential und einer effektiveren Induktion einer Th1-Cytokin-Antwort äußerte. Unter serumfreien Bedingungen generierte unreife DC zeigten eine typische potente Antigenaufnahmeaktivität, welche für die Beladung mit Tumorantigenen erforderlich ist. Reife DC zeigten u.a. eine charakteristische Morphologie und eine T-Zell-stimulatorische-Aktivität. Wurden serumfrei generierte DC mit Tumorantigen der Patienten beladen, so induzierten sie eine Tumorantigen-spezifische autologe T-Zell-Antwort. Da Tumoren während ihrer Entwicklung kontinuierlich immunologisch selektioniert werden, wurde abschließend untersucht, ob Gliomzellen mit der durch DC induzierten Immunität interferieren. Hierbei zeigte sich, dass die Gliomzellen Faktoren sezernieren, welche die Differenzierung und Ausreifung DC hemmen. So war die Expression der immunrelevanten Oberflächenmoleküle CD40, CD80,CD86, HLA-Klasse I/II sowie von sezernierten Cytokinen wie IL-12 und von zahlreichen Chemokinen supprimiert, was sich funktionell in einer reduzierten T-Zell-stimulatorischen Aktivität der DC und einer verminderten Produktion des Th1-Cytokins IFNgamma durch die stimulierten T-Zellen äußerte. Als Kandidaten für diese immunsupressive Aktivität der Gliomzellen wurde VEGF, IL-6 und TGFbeta nachgewiesen, doch ihre Neutralisierung konnte die Differenzierung und Ausreifung der DC nur partiell wiederherstellen.
... SVV presents during fetal development, but undetectable in terminally differentiated adult tissues [5]. The survivin gene locus encodes multiple genetic splice variants with unique properties and functions [6] [7]. ...
... Wild type survivin expression increased in stage III (p = 0.05) and stage IV (p = 0.02) as compared to stage I, indicating a direct relationship between survivin gene expression and disease progression in breast cancer. Survivin-ΔEx3 overexpression has been associated with high proliferation of tumor cells (Yamada et al., 2003) and poor prognosis in cancer (Takashima et al., 2005;Ling et al., 2005). In our study, we found that survivin-ΔEx3 gene expression was significantly higher in advanced stages of cancer i.e., stages II-III as compared to stage I (Taubert et al., 2005a). ...
... Interestingly, expression of other human SVV isoforms such as SVV-2B and SVV-deltaEx3 were mostly reported in cancer patients, in which the expressions were upregulated at different stages of tumor development [33][34][35][36]. Under normal physiology, however, SVV-α is the main isoform identified in humans, which is known to regulate cell mitosis [32,37]. This concurs with the finding of SVV in the nucleus but not in the cytosol of cardiomyocytes during embryonic heart development, which indicates that the presence of SVV in the nucleus was critical for cell mitosis to happen in embryonic hearts. ...
Rationale:Reducing cardiomyocyte death and enhancing their proliferation after myocardial infarction is perhaps the single largest challenge for cardiac tissue regeneration. Survivin (SVV) is the smallest member of the inhibitor of apoptosis (IAP) family but plays two important roles; inhibiting caspase-9 activation in the intrinsic apoptosis pathway, and regulating microtubule dynamics and chromosome segregation during cell division. Genetic depletion of cardiac SVV leads to incomplete cardiomyocyte division and abnormal heart development. However, the function of SVV in adult hearts after myocardial infarction remains unclear.
Methods: A homozygous inducible cardiomyocyte-specific SVV knockout transgenic mouse model was established through crossbreeding SVVflox/flox and αMHC-MCM transgenic mice. Adult mice received consecutive intraperitoneal injection of tamoxifen to induce genetic removal of SVV in cardiomyocytes. A SVV overexpressing model was established via local delivery of SVV in wild-type mouse hearts.
Results: We found that 30.82% of cardiomyocytes in the peri-infarct region of SVV knockout mice were apoptotic, significantly higher than the 22.18% in control mice. In addition, ejection fraction was 29.00±0.40% in knockout mice compared to 38.04±0.50% in control mice 21 days after myocardial infarction. On the contrary, locally overexpressing SVV in the heart improved cardiac functions. Unexpectedly, we found that altering the subcellular localization of SVV overexpression produced different outcomes. Overexpression of SVV in the cytoplasm decreased cardiomyocyte apoptosis, whereas overexpression of SVV in the nucleus enhanced cardiac regeneration. The ejection fraction of mice overexpressing SVV was 36.58±0.91%, significantly higher than 28.18±1.70% in the GFP control group. Apoptotic cardiomyocytes were only 4.63% in mouse overexpressing cytosolic SVV, compared to 9.31% in the GFP group, and activation of caspase-3 was also reduced. Moreover, mice overexpressing NLS-SVV exhibited a better ejection fraction (36.19±1.02%,) than GFP controls (26.69±0.75%). NLS-SVV enhanced H3P-positive cardiomyocytes in the border zone to 0.28%, compared to only 0.08% in GFP group, through interacting with Aurora B.
Conclusions:We demonstrate the importance of SVV subcellular localization in regulating post-MI cardiac repair and regeneration. We hope that this will open new translational approaches through targeted delivery of SVV.
... Moreover, although Krieg et al. indicated that survivin and survivin-4Ex3 remained unchanged in different stages of cancer, Meng et al. showed that the expression level of survivin-4Ex3 was inversely correlated with the apoptotic index [39]. In addition, survivin-4Ex3 and survivin-2B may play opposing roles in tumor progression and/or tumorigenesis [40]. In our meta-analysis, the antibodies used in most studies do not discriminate between survivin isoforms, and thus the results may reflect increases in total survivin levels, including all known isoforms. ...
Objective
Overexpression of survivin has been reported in many human tumors. However, the clinicopathological features associated with survivin overexpression in cervical carcinoma remain controversial. Thus, the current meta-analysis was performed to assess the clinicopathological significance of survivin in cervical carcinoma.
Methods
PubMed, EMBASE, and Web of Science databases were searched for relevant studies published through November 1, 2015. A meta-analysis was performed to evaluate the association between survivin expression and clinicopathological outcome in cervical carcinoma.
Results
Eleven eligible studies with a total of 865 patients were included. Survivin overexpression was closely related to lymph node metastasis (odds ratio [OR] = 0.679, 95% confidence interval [CI]: 0.509–0.905, P = 0.008) but was not significantly associated with tumor FIGO stage (I+II vs. III+IV) (OR = 0.843, 95% CI: 0.626–1.137, P = 0.264), tumor grade (G1+G2 vs. G3) (OR = 0.913, 95% CI: 0.689–1.210, P = 0.527), tumor size (>4 vs. ≤4 cm) (OR = 0.825, 95% CI: 0.434–1.570, P = 0.559), or stromal involvement (OR = 0.820, 95% CI: 0.545–1.233, P = 0.340). The correlation between survivin expression and overall survival was evaluated among a total of 238 patients from three eligible studies. The pooled HR was 1.129 (95% CI: 0.597–1.661; P = 0.000), indicating that survivin expression was significantly associated with poor survival in cervical carcinoma.
Conclusions
Based on the current meta-analysis, survivin is strongly associated with lymph node metastasis and poor prognosis. Additionally, survivin is a novel clinicopathological marker of cervical carcinoma and thus may be a therapeutic target for cervical carcinoma.
... The median survival among patients with GBM was 622 days. Other tumor vaccine targets under investigation include cytochrome p450, [23] telomerase, [24] GALT3 ( b GlcNAc b 1, 3-galactosyltransferase, polypeptide 3), [25] survivin, [26] tenascin, [27] glycoprotein 240, [28] and SART AQ16 ((Author: please define SART)) 1. [29] ...
Malignant gliomas represent one of the most aggressive forms of brain cancer. Recent advances in the understanding of the deregulated molecular pathways of gliomas have brought about targeted therapies that have the ability to increase therapeutic efficacy in tumors while decreasing toxicity. Multi-targeted kinase inhibitors, novel monoclonal antibodies, and new vaccines have been developed. Standard treatments and current development of new therapies for malignant gliomas are reviewed, focusing specifically on growth factors and their receptors (e.g. epidermal growth factor receptor, vascular endothelial growth factor receptor, and platelet-derived growth factor receptor), as well as the intracellular effector molecules that are downstream of these growth factors (e.g. Ras/Raf/mitogen-activated protein kinase, phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin, and protein kinase C). The efficacies of other novel targeted inhibitors such as deacetylase inhibitors and heat shock protein 90 inhibitors in the treatment of gliomas are also discussed, as well as new combination therapies. In order for new agents to increase treatment efficacy, new targets need to be developed, drug delivery efficiency needs to be improved, and new biomarkers need to be discovered. All of these goals can be accomplished with time through innovative experimental designs.
... Apoptosis is modulated by a number of gene products which act through protein-protein interactions either as inducers or inhibitors. In our study, the genes of DAP, PHLDA, ATF 2 , p21waf 1 /cip 1 , and their products which could induce cellular apoptosis [22][23][24][25][26] , were obviously up-regulated in SLHCC compared with NHCC, whereas survivin, which inhibited cellular apoptosis [27][28][29][30][31] was less in SLHCC than in NHCC. These data suggested that one of the mechanisms of SLHCC, which possesses a less invasive and metastatic potential than NHCC, might be a facility in apoptosis of HCC cells. ...
AIM: To study the difference in gene expression between solitary large hepatocellular carcinoma (SLHCC) and nodular hepatocellular carcinoma (NHCC).
METHODS: Polymerase chain reaction (PCR) products of 8464 human genes were spotted on a chip in array. DNAs were then fixed on a glass plate. Total RNA was isolated from freshly excised human SLHCC (n = 7) and NHCC (n = 15) tissues, and was reversely transcribed to cDNAs with the incorporation of fluorescent dUTP for preparation of hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After highly stringent washing, cDNA microarray was scanned for the fluorescent signals to display the difference between the two kinds of HCC. In addition, the expression of RhoC and protocadherin LKC was also detected with the reverse transcriptase polymerase chain reaction (RT-PCR) method.
RESULTS: Among the 8464 human genes, 668 (7.89%) genes were expressed differentially at the mRNA levels between SLHCC and NHCC. Three hundred and fifty five (4.19%) genes, including protocadherin LKC, were up-regulated, whereas 313 (3.70%) genes, including RhoC, were down-regulated. The mRNA expression levels of RhoC and protocadherin LKC were confirmed by RT-PCR. Analysis of differentially expressed genes confirmed that our molecular data obtained by cDNA microarray were consistent with the published biochemical and clinical observations of SLHCC and NHCC.
CONCLUSION: cDNA microarray is an effective technique in screening the difference in gene expression between SLHCC and NHCC. Many of these differentially expressed genes are involved in the invasion and metastasis of HCC. Further analysis of these genes will help to understand the different molecular mechanisms of SLHCC and NHCC.
Keywords: $[Keywords]
Citation: Yang LY, Wang W, Peng JX, Yang JQ, Huang GW. Differentially expressed genes between solitary large hepatocellular carcinoma and nodular hepatocellular carcinoma. World J Gastroenterol 2004; 10(24): 3569-3573
... Quantifying the levels of Survivin and its splice variants is useful for the prediction of the cell biological malignancy of glioma, independent of their pathological features (Yamada, Kuroiwa et al. 2003). It is also useful for demonstrating tumours in which a tricyclic antidepressant might be a useful therapy. ...
... This approach has already been tested by several clinical trials [27]. Many other HGG proteins such as EphA2 (Ephrin type-A receptor 2) [28] , IL-13 receptor α2 chain [29] , surviving [30], tenascin [31], and SOX2 [32] might serve as markers for gliomas. These studies need further exploration due to the limited sample size and heterogeneous expression level of glioma-specific proteins. ...
The prognosis of patients with high grade gliomas (HGG) is quite poor, in spite of the standard combination of surgery, radiotherapy and chemotherapy. Recently, many clinical researchers have been focusing their efforts on the safety, feasibility and efficiency of dendritic cells (DCs) based vaccine against HGG. According to the literature search, 23 phase I/II clinic studies were picked and systemically reviewed to assess the application, the effect, and the future of DC vaccine for HGG. DC vaccine appears to have potential to increase overall survival with minimal complications. However, there still remains to be many challenges during vaccination, such as targeting specific/ associated antigens, adjuvants, clinical status, and the evaluation of the response.
... This study is the first examination of the expression of survivin splice variants in benign tumors and indicated that the expression of survivin in benign tumors was significantly higher compare with malignant ones. However, several studies have reported the relevance of survivin expression with malignant progression (Yamada et al., 2003;Zhang et al., 2004;Ryan et al., 2005;Sah et al., 2006). The expression of survivin-DEx3 was detected in 24 out of 25 (95%) malignant tumors, while it was expressed weakly in only a few benign tumors. ...
Survivin, which is a novel member of the inhibitor of apoptosis family proteins, is known to play an important role in the regulation of cell cycle and apoptosis. Differential expression of survivin in tumor tissues introduces it as a new candidate molecular marker for cancer. Here we investigated the expression of survivin and its splice variants in breast tumors, as well as normal adjacent tissues obtained from the same patients. Thirty five tumors and 17 normal adjacent tissues from women diagnosed with breast cancer were explored in this study. Differential expression of different survivin splice variants was detected and semiquantitatively analyzed using reverse transcription–polymerase chain reaction. Results showed that survivin and its splice variants were differentially expressed in tumor specimens compared with normal adjacent tissues. The expression of survivin-3B and survivin-3a was specifically detected in tumor tissues compared with normal adjacent ones (53% in tumor tissues compared to 5% in normal adjacent for survivin-3B and 65% in tumor tissues and 0.0% in normal adjacent tissues for survivin-3a). Statistical analysis showed that survivin and survivin-DEx3 were upregulated in benign (90%, p < 0.034) and malignant (76%, p < 0.042) tumors, respectively. On the other hand, our results showed that survivin-2a (100% of the cases) was the dominant expressed variant of survivin in breast cancer. The data presented here showed that survivin splice variants were differentially expressed in benign and malignant breast cancer tissues, suggesting their potential role in breast cancer development. Differential expression of survivin-2a and survivin-3a splice variants highlights their usefulness as new candidate markers for breast cancer diagnosis and prognosis.
... Pituitary adenomas are the most common tumors in the central nervous system and are thought to be monoclonal in origin [1]. ...
Purpose:
Survivin is an apoptosis inhibitor, expressed in almost all types of human malignancies, but rarely in differentiated normal tissues. Recently, survivin gene splice variants with different anti-apoptotic activities have been reported. The current study was undertaken to examine the expression of survivin and its splice variants ∆Ex3 and 2β in pituitary tumors, and to correlate the amount of particular transcripts with clinical staging in pituitary adenomas. Quantitative detection of survivin and its splice variants ∆Ex3 and 2β transcripts in non-cancerous pituitary tissues (n = 12) and different types of pituitary tumor (n = 50).
Methods:
Samples were collected from 50 pituitary tumors including 26 non-functional tumors, 21 GH-secreting tumors, 2 PRL-secreting tumors and 1 ACTH-secreting tumor. 12 normal pituitary glands received after autopsy served as a control of the study. 29 thyroid cancers tissues were used as a positive control. The RT-qPCR with TaqMan hydrolysis probes were used to determine the expression of analyzed splice variants of survivin.
Results:
The obtained data showed that both survivin and its splice variants were expressed in different types of pituitary adenoma as well as in normal pituitary tissue. However, the level of its expression was similar in all studied cases. Patient age negatively correlated with tumor invasiveness. Moreover, our study showed a tendency for negative correlation between patient age and tumor diameter.
Conclusions:
No significant differences between survivin and its splice variants ∆Ex3 and 2β expression in pituitary tumors and in normal pituitary glands as well as in invasive and in non-invasive tumors were found, suggesting that survivin does not play a significant role in pituitary tumorigenesis.
... 7 A previous report showed that expression of both survivin variants 1 and 2 was significantly higher in malignant brain tumors relative to benign ones (this correlation did not hold for variant 3). 30 A study using nonquantitative RT-PCR techniques showed that survivin variants 1 and 2 were detectable in a significant number of glioblastoma cases and had a strong negative correlation with survival in cases displaying mRNA positivity for variants 1 and 2. 8 Our analysis of the TCGA dataset was calibrated to determine how expression above normalized mean expression of these variants correlated with survival. Therefore, our analysis shows that variant 2 is less frequently overexpressed in human glioma. ...
The influence of survivin isoforms on outcome in glioblastoma is poorly understood. We analyzed the dominant anti-apoptotic transcript variants of survivin using expression data and modeled them in vivo to determine their impact on glioma formation and progression.
Using data from low- and high-grade glioma knowledge bases, we expressed the anti-apoptotic isoforms of survivin (transcript variants 1 and 2) in vivo using the RCAS/Ntv-a model of murine glioma.
In low-grade gliomas, survivin RNA expression was increased in 22 of 167 (13.2%) of cases and was associated with shortened survival (P = .005). Survivin RNA was preferentially expressed in proneural (PN) relative to mesenchymal high-grade gliomas (P < .0001). In proneural gliomas, survivin was expressed in 94 of 141 (67%) of cases and was associated with shorter disease-free survival (P = .04). In a platelet-derived growth factor subunit B-dependent murine model of PN glioma, ectopic expression of variant 1 yielded tumors in 28 of 30 (93%) of mice, of which 25% were high-grade tumors, whereas ectopic expression of variant 2 yielded tumors in 27 of 28 (96%), of which 81% were high-grade tumors (P < .0001). Microvascular proliferation was significantly more prominent (P < .0001), and tumor-free survival was shorter in mice with variant 2 than variant 1-derived tumors (P = .01).
Survivin expression in low-grade gliomas is associated with poor survival and is preferentially expressed in PN gliomas. Compared with variant 1, variant 2 was associated with poorer survival and promoted malignant progression, angiogenesis, and shorter tumor-free survival in the PN murine model. Inhibiting survivin transcript variant 2, rather than variant 1 (the common isoform), may be an effective treatment strategy for glioma.
... This study is the first examination of the expression of survivin splice variants in benign tumors and indicated that the expression of survivin in benign tumors was significantly higher compare with malignant ones. However, several studies have reported the relevance of survivin expression with malignant progression (Yamada et al., 2003;Zhang et al., 2004;Ryan et al., 2005;Sah et al., 2006). The expression of survivin-DEx3 was detected in 24 out of 25 (95%) malignant tumors, while it was expressed weakly in only a few benign tumors. ...
Survivin, which is a novel member of the inhibitor of apoptosis family proteins, is known to play an important role in the regulation of cell cycle and apoptosis. Differential expression of survivin in tumor tissues introduces it as a new candidate molecular marker for cancer. Here we investigated the expression of survivin and its splice variants in breast tumors, as well as normal adjacent tissues obtained from the same patients. Thirty five tumors and 17 normal adjacent tissues from women diagnosed with breast cancer were explored in this study. Differential expression of different survivin splice variants was detected and semiquantitatively analyzed using reverse transcription-polymerase chain reaction. Results showed that survivin and its splice variants were differentially expressed in tumor specimens compared with normal adjacent tissues. The expression of survivin-3B and survivin-3α was specifically detected in tumor tissues compared with normal adjacent ones (53% in tumor tissues compared to 5% in normal adjacent for survivin-3B and 65% in tumor tissues and 0.0% in normal adjacent tissues for survivin-3α). Statistical analysis showed that survivin and survivin-ΔEx3 were upregulated in benign (90%, p<0.034) and malignant (76%, p<0.042) tumors, respectively. On the other hand, our results showed that survivin-2α (100% of the cases) was the dominant expressed variant of survivin in breast cancer. The data presented here showed that survivin splice variants were differentially expressed in benign and malignant breast cancer tissues, suggesting their potential role in breast cancer development. Differential expression of survivin-2α and survivin-3α splice variants highlights their usefulness as new candidate markers for breast cancer diagnosis and prognosis.
... On the basis of our proposed concept we would predict that Sox2 and/or survivin would be highly expressed in brain tumours. This prediction agrees with current findings as reported by Yamada et al. [27] that abnormal expression of Sox2 or survivin is associated with brain tumour formation [28]. Furthermore, we would expect to observe a loss in NSCs in the brains of aging and neurological degenerative patients, which may be associated with a deficiency in Sox2 and/or survivin production. ...
Transcriptional factor Sox2 is essential for regulation of self-renewal and homeostasis of neural stem cells (NSCs) during brain development. However, the downstream targets of Sox2 and the underlying molecular mechanism are largely unknown. In this study, we found that Sox2 directly up-regulates the expression of survivin, which inhibits the mitochondria-dependent apoptotic pathway in NSCs. While over-expression of Sox2 elevates the survivin expression, knock-down of Sox2 results in a decrease in survivin expression, thereby initiating the mitochondria-dependent apoptosis related with caspase-9 activation. Furthermore, cell apoptosis due to Sox2 knock-down can be rescued by ectopically expressing survivin in NSCs as well as in the mouse brain, as demonstrated by an in utero-injection approach. In short, we have found a novel Sox2-survivin pathway that regulates NSC survival and homeostasis, thus revealing a new mechanism of brain development, neurological degeneration and such aging-related disorders.
... Median survival among GBM patients was 622 days. Other tumor vaccine targets under investigation include cytochrome p450 [184], telomerase [185], GALT3 [186], survivin [187], tenascin [188], glycoprotein 240 [189], and SART1 [190]. ...
Battling aggressive brain tumors represents a struggle endeavored by scientists and physicians across many different disciplines.
The molecular and physiological abnormalities that accompany this fatal disease render malignant brain tumors arguably one
of the most difficult pathologies to treat. For this reason, the efforts to combat this fatal disease are represented by a
plethora of research. This chapter is organized to illustrate common themes in drug targeting for the treatment of brain tumors.
Each section is organized based on the therapeutic approach: small molecules, local drug delivery, or biological therapy.
In each subsection we discuss the rationale that has ushered each development to the ‘center stage’ in brain tumor therapy
and provide illustrative examples using the most relevant and heretofore advanced developments. In this way, we hope to provide
a discussion of brain tumor therapy that is all-encompassing yet detailed enough to give the reader a broad understanding
of how researchers and clinicians attempt to improve the outcome of patients diagnosed with this debilitating disease.
... Suga et al. [45] found that colorectal carcinoma patients having a higher mRNA expression ratio of survivin-2b/wild-type survivin were associated with better prognosis and in early-stage disease, while patients with a lower expression of this ratio had shorter survival and advanced stage disease. Regarding the cellular level of survivin-2b, its expression was higher in less aggressive 24 15 neuroblastoma and benign brain tumors when compared to their corresponding more aggressive types [46,47]. Furthermore, Mahotka et al. [48] also observed a significant decrease in survivin-2b level during the progression of renal cell carcinoma. ...
To examine the correlation of survivin (both total and nuclear survivin) with clinicopathological parameters of esophageal squamous cell carcinoma (ESCC) patients. Tumors and non-tumor tissues near the proximal resection margins were resected from ESCC patients undergone esophagectomy. Quantitative polymerase chain reaction (qPCR) was performed to detect survivin mRNA expression level in the 10 paired tumor and adjacent non-tumor tissues. To confirm with the clinical situation, survivin mRNA and protein expression were measured by qPCR and immunoblot, respectively, in 5 ESCC cell lines and a non-neoplastic esophageal epithelial cell line. Immunohistochemistry was employed to reveal the cellular localization of survivin in tumor tissues isolated from the 64 ESCC patients undergone surgery alone. Up-regulation of survivin mRNA and protein was found in 5 ESCC lines (HKESC-1, HKESC-2, HKESC-3, HKESC-4, and SLMT-1) when compared to a non-neoplastic esophageal epithelial cell line NE-1. In particular, HKESC-3, HKESC-4, and SLMT-1 cells demonstrated ~50-fold increase in survivin mRNA. High level of survivin mRNA in tumor tissues when compared to non-tumor tissues was found in 70 % (7 of 10) of clinical cases. The increase in expression ranged from ~twofold to ~16-fold. Immunohistochemistry results showed that survivin was found at the cell nuclei in all specimens examined. Nuclear expression of survivin was inversely associated with the likelihood of developing nodal metastasis (p = 0.021) and significantly associated with early-stage ESCC (p = 0.039). Nuclear survivin could serve as a marker for indicating disease status in ESCC patients.
... Hypoxia induces MDK expression through the binding of to a hypoxia responsive element in the MDK promoter. Survivin, an antiapoptotic protein, has been found to be overexpressed in up to 79% of astrocytic tumors (Kajiwara et al., 2003; Yamada et al., 2003; Chakravarti et al., 2002). The expression of this gene correlates with grade and is present in 90% of GBMs.The activity of this promoter is also enhanced by hypoxia, commonly found in rapidly growing tumors like high grade gliomas (Yang et al., 2004). ...
... However, there have been reports that the expression of survivin splice variants significantly correlates with the clinical results in many types of human carcinoma, including neuroblastoma, renal cell carcinoma, gastric cancer, brain tumor, leukemia, and breast cancer. Many investigators have suggested that sur- vivin-ΔEx3 and survivin-2B play opposing roles in tumor progression and/or tumorigenesis789101112. The over-expression of survivin in colorectal carcinoma has been identified as an independent prognostic factor [13,14] . ...
Recently, two alternatively spliced survivin variants, survivin-ΔEx3 and survivin-2B, were identified in a single copy of the survivin gene. It has been reported that the expressions of survivin splice variants significantly correlates with the clinical results in many types of human carcinoma. We investigated the transcription levels of survivin and its splice variants in human colorectal carcinomas, and analyzed correlations between survivin expression levels and clinicopathologic features.
We used Western blot and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) to analyze the protein and mRNA expression levels of survivin variants in 51 colorectal carcinomas. The quantitative RT-PCR was performed using primer pairs specific for survivin and each of its splice variants, then normalized for the gene that encodes glyceraldehydes-3-phosphate dehydrogenase.
In Western blotting, the protein levels of survivin were higher in the tumor tissue than in normal tissue. The expression of survivin, survivin-2B and survivin-ΔEx3 mRNA was present in 96%, 64.7%, and 82.4% of the samples, respectively. When the pathologic parameters were compared, colorectal cancers of advanced pT stages showed significant decrease in survivin-2B mRNA expression by the quantitative RT-PCR (P < 0.001).
The decreased expression of survivin-2B might be related to tumor progression in colorectal cancers. This finding indicates that alternatively spliced variants of survivin may be involved in refining the functions of survivin during tumor progression.
... [21] Anti apoptotic properties of survivin enhance the viability of cancer cells. [13] It has been reported that the survivin and its splice variants are highly expressed in several human carcinoma including lung, breast, colon and oesophageal compared with normal tissues.22232425262728 Moreover Ito et al., reported the high expression level of survivin in human thyroid tumors using immunohistochemistry technique. ...
The most important problem in the case of thyroid nodules is the lack of suitable criteria for detecting malignant thyroid tumors from other nodules in the early stage. Variable expressions level of survivin, an inhibitory protein in apoptotic pathway, and its splice variants in malignant carcinoma versus well-differentiated normal tissues candidate them as reliable biomarkers in cancers.
To semi-quantitative detection of survivin and its splice variant, survivin-deltaEx3, in thyroid nodules.
We evaluated the expression level of mentioned biomarkers in thyroid nodules including carcinoma.
Samples were collected from 61 thyroid nodules including malignant, adenoma, non-tumoral (goiter and thyroidities) as well as non-neoplastic normal tissues. Transcriptional levels were measured using semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and the results were normalized to β2microglubin (β2m) gene. Statistical Analysis Used: Independent sample t-test was used to assess the significant variation of expression between different groups.
Our data for a first time revealed that survivin-deltaEx3 is significantly up-regulated from normal to malignant thyroid carcinoma tissues (approximately ten fold).
High expression level of survivin and survivin-deltaEx3 in malignant papillary thyroid carcinoma suggested survivin gene expression and its splice variant, survivin-deltaEx3, can be potential new markers in diagnosis of human papillary thyroid carcinoma.
... Given the complexity of splicing regulation in the brain, aberrant splicing in gliomas may be a significant but yet under-explored contributor to the heterogeneous pathological characteristics of GBM. Indeed, glioma-specific AS patterns (Cheung et al, 2008) have been reported, as well as additional aberrant splicing events that can contribute to all aspects of gliomagenesis , affecting angiogenesis (Huang et al, 2005 ), oncogenic suppression (Chunduru et al, 2002), escape from apoptosis (Yamada et al, 2003), proliferation (CamachoVanegas et al, 2007; Yu et al, 2007), metabolism (Clower et al, 2010; David et al, 2010) and migration/invasion (Yu et al, 2007; Cheung et al, 2009; Lo et al, 2009). understand the role of AS deregulation in GBM, we looked at previously described splicing events that are altered in a variety of cancers and can generate variants with experimentally verified oncogenic properties. ...
In tumours, aberrant splicing generates variants that contribute to multiple aspects of tumour establishment, progression and maintenance. We show that in glioblastoma multiforme (GBM) specimens, death-domain adaptor protein Insuloma-Glucagonoma protein 20 (IG20) is consistently aberrantly spliced to generate an antagonist, anti-apoptotic isoform (MAP-kinase activating death domain protein, MADD), which effectively redirects TNF-α/TRAIL-induced death signalling to promote survival and proliferation instead of triggering apoptosis. Splicing factor hnRNPH, which is upregulated in gliomas, controls this splicing event and similarly mediates switching to a ligand-independent, constitutively active Recepteur d'Origine Nantais (RON) tyrosine kinase receptor variant that promotes migration and invasion. The increased cell death and the reduced invasiveness caused by hnRNPH ablation can be rescued by the targeted downregulation of IG20/MADD exon 16- or RON exon 11-containing variants, respectively, using isoform-specific knockdown or splicing redirection approaches. Thus, hnRNPH activity appears to be involved in the pathogenesis and progression of malignant gliomas as the centre of a splicing oncogenic switch, which might reflect reactivation of stem cell patterns and mediates multiple key aspects of aggressive tumour behaviour, including evasion from apoptosis and invasiveness.
... Studies using sensitive quantitative Real time-PCR revealed that survivin mRNA was expressed in normal fetal tissue panels and at a very low level in the normal adult brain template. In contrast, data from a recent study have demonstrated survivin expression in normal adult tissues, such as those of skin, endometrium, endothelial cells, normal blood lymphocytes, pancreas, spleen, colon, stomach, small intestine, large intestine, lung, kidney, prostate, pancreas, heart, and thymus [45] . By using sensitive quantitative Real time-PCR analysis, low levels of total survivin mRNA were detectable in normal tissue adjacent to soft tissue sarcoma tumor cells (e.g. ...
To investigate three isoforms of survivin in colorectal adenocarcinomas.
We used the LightCycler Technology (Roche), along with a common forward primer and reverse primers specific for the splice variants and two common hybridization probes labeled with fluorescein and LightCycler-Red fluorophore (LC-Red 640). Real time quantitative polymerase chain reaction (PCR) was performed on cDNAs from 52 tumor specimens from colorectal cancer patients and 10 unrelated normal colorectal tissues. In the patients group, carcinoembryonic antigen (CEA) and CA19-9 tumor markers were also measured immunochemically.
Wild type survivin mRNA isoform was expressed in 48% of the 52 tumor samples, survivin-2b in 38% and survivin-ΔΕx3 in 29%, while no expression was found in normal tissues. The mRNA expression of wild type survivin presented a significant correlation with the expression of the ratio of survivin-2b, survivin-ΔΕx3, survivin-2b/wild type survivin and survivin-ΔΕx3/wild type survivin (P < 0.001). The mRNA expression of wild-survivin and survivin-ΔΕx3 was related with tumor size and invasion (P = 0.006 and P < 0.005, respectively). A significant difference was found between survivin-2b and morphologic cancer type. Also, the ratio of survivin-ΔEx3/wild-survivin was significantly associated with prognosis. No association was observed between the three isoforms and grade, metastasis, Dukes stage and gender. The three isoforms were not correlated with CEA and CA19-9.
Survivin isoforms may play a role in cell apoptosis and their quantification could provide information about clinical management of patients suffering from colorectal cancer.
... Die gezielte Expression von Survivin-2B oder die Hemmung von Survivin-∆3 könnte als Tumortherapie Anwendung finden. Auch in normalem Hirngewebe konnten Survivin und seine Spleißvarianten nachgewiesen werden, wobei das Verhältnis der Expression von Survivin-∆3/Survivin signifikant höher in malignen als in benignen Hirntumoren war(Yamada et al. 2003, Altura et al. 2003.Eine andere Möglichkeit, die Survivin-exprimierenden Tumorzellen zu schädigen, ist der Einsatz immunologischer Verfahrensweisen. Immunzellen (CD8+ zytotoxische T-Lymphozyten) erkennen an der Zelloberfläche von Tumorzellen präsentierte Survivinproteinfragmente(Rohayem et al. 2000, Andersen et al. 2001, Hirohashi et al. 2002. ...
Halle, Wittenberg, Univ., Diss., 2005. Computerdatei im Fernzugriff.
... InteressanteVarianten des IAP (inhibitor of apoptosis) Survivin (Ambrosini et al. 1997) variieren in normalen und Tumorzellen, und dienen als ein Indikator für maligne Tumoren (Yamada et al. 2003). Erst kürzlich konnte gezeigt werden, dass eine weitere Spleiß-Variante von Survivin, Survivin 3B, in einigen Tumoren exprimiert wird und dort evtl. ...
Charakterisierung eines zuvor funktionell unbeschriebenen Genes, das in einem genetischen Screen nach proapoptotischen Genen isoliert wurde und RAIP genannt wurde. Verifizierung der proapoptotischen Eigenschaften mit mehreren Apoptose-Assays in humanen Zelllinien, Nachweis der Lokalisation im ER in Kulturzellen, Eingrenzung eines 63 Aminosäure-Reste grossen proapoptotischen Fragmentes, Isolierung von drei Interaktionspartnern (Ferritin, SRp40, SIRTUIN 7).
Background:
Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease with poor prognosis, as it is difficult to predict or circumvent, and it develops chemoresistance quickly. One cellular mechanism associated with chemoresistance is alternative splicing dysfunction, a process through which nascent mRNA is spliced into different isoforms. Survivin (Baculoviral IAP Repeat-Containing Protein 5 (BIRC5)), a member of the inhibitor of apoptosis (IAP) protein family and a cell cycle-associated oncoprotein, is overexpressed in most cancers and undergoes alternative splicing (AS) to generate six different splicing isoforms.
Methods:
To determine if survivin splice variants (SSV) could be involved in PDAC chemoresistance, a Gemcitabine (Gem) resistant (GR) cell line, MIA PaCa-2 GR, was created and assessed for its SSV levels and their potential association with GR. Cross-resistance was assessed in MIA-PaCa-2 GR cells to FIRINOX (5-fluorouracil (5-FU), irinotecan, and oxaliplatin). Once chemoresistance was confirmed, RT-qPCR was used to assess the expression of survivin splice variants (SSVs) in PDAC cell lines. To confirm the effect of SSVs on chemoresistance, we used siRNA to knockdown all SSVs or SSV 2β.
Results:
The MIA PaCa-2 GR cell line was 40 times more resistant to Gem and revealed increased resistance to FIRINOX (5-fluorouracil (5-FU), irinotecan, and oxaliplatin); when compared to the parental MIA-PaCa-2 cells. RT-qPCR studies revealed an 8-fold relative expression increase in SSV 2β and a 2- to 8-fold increase in the other five SSVs in the GR cells. Knockdown of all SSV or SSV 2β only, using small inhibitory RNA (siRNA), sensitized the GR cells to Gem, indicating that these SSVs play a role in PDAC chemoresistance.
Conclusion:
These findings provide evidence for the potential role of SSV 2β and other SSVs in innate and acquired PDAC chemoresistance. We also show that the expression of SSVs is not affected by the type of chemoresistance, therefore targeting survivin splice variants in combination with chemotherapy could benefit a wide range of patients.
OBJECTIVE
The object of this study was to clarify the expression characteristics and prognostic value of survivin in skull base chordomas.
METHODS
In this retrospective study, the authors measured the expression of survivin at the mRNA level in 81 samples from 71 patients diagnosed with skull base chordomas at their hospital in the period from July 2005 to January 2015. Clinical data collection, follow-up, and survival analyses were performed, and correlations were analyzed.
RESULTS
Of the 71 patients, 50 had primary chordomas with a mean survivin expression level of 1.09; the other 21 patients had recurrent chordomas with a mean survivin expression level of 2.57, which was 2.36 times higher than the level in the primary chordoma patients (p < 0.001, Mann-Whitney U-test). In addition, an analysis of 18 paired samples derived from 9 patients showed that the expression level of survivin was 2.62 times higher in recurrent tumors than in primary tumors (p = 0.002, paired t-test). The Spearman rank correlation coefficient method showed that the expression level of survivin was positively correlated with the mean ratio of tumor signal intensity to the signal intensity of surrounding brainstem on T1-weighted sequences (R T1 ; r s = 0.274, p = 0.021) and was negatively correlated with the mean ratio of tumor signal intensity to the signal intensity of surrounding brainstem on T2-weighted sequences (R T2 ; r s = −0.389, p = 0.001). A multivariate Cox proportional-hazards model suggested that pathology (p = 0.041), survivin expression level (p = 0.018), preoperative Karnofsky Performance Status (KPS; p = 0.012), and treatment history (p = 0.009) were independent prognostic factors for tumor progression. Survivin expression level (p = 0.008), preoperative KPS (p = 0.019), tumor diameter (p = 0.027), and intraoperative blood loss (p = 0.015) were independent prognostic factors for death.
CONCLUSIONS
Survivin expression level and preoperative KPS were independent significant prognostic factors for tumor progression and death in skull base chordoma patients. Recurrent skull base chordomas and chordomas with high R T1 and low R T2 were likely to have high survivin expression. Other independent risk factors related to tumor progression included conventional pathology and treatment history, whereas additional mortality-related risk factors included larger tumor diameter and greater intraoperative blood loss.
Chapter 8 addresses three different tumors: medulloblastoma, primitive neuroectodermal tumors, and pineal tumors. The genetic and molecular studies in medulloblastoma are legion; yet, the basic changes responsible for the development of these tumors have remained evasive. Nevertheless, cytogenetically about one third of the medulloblastomas contain loss of 1p or i(17q), and less common anomalies of chromosomes 1 and 7, loss of chromosome 7, and loss of chromosome 22 (-22) and other changes. Of interest is the presence of double minutes in some medulloblastomas (approx 7%). The aforementioned changes have been supplemented with those obtained by spectral karyotyping, fluorescence in situ hybridization, comparative genomic hybridization (CGH), and loss of heterozygosity (LOH) results. Possible candidate genes playing a role in medulloblastoma development may reside at 10q or 8p. The changes of MYCN (amplification) in approximately 10% of medulloblastomas are of interest. Other genes investigated in these tumors include TP53 and ARF, IGF, epidermal growth factor receptor (EGFR)-ErbB, PDFGFα, RAS/MAPK, and PAX and ENI. The sonic hedgehog (SHH) and Wingless signaling pathways may play a role in medulloblastoma biology, and also neurotrophins and other factors (related to apoptosis), such as somatostatin receptors, Notch signaling, epigenetic events, the HIC gene, caspase8 gene, RASSF1A gene, and telomerase activity. The hereditary and genetic susceptibilities to medulloblastoma development are discussed. Chapter 8 also addresses primitive neuroectodermal tumors (PNET) cytogenetically; the changes in PNET show a variable pattern, as do the changes obtained with CGH and LOH. Molecular studies of PNET have included p53, Notch signaling, and the SHH pathway. Approaches similar to those used for the study of PNET have been applied to pineal tumors with the results being variable and not specific for any of the tumors.
Purpose:
Gliomas are the most common primary brain tumors. The etiology is still unclear and the progression from low to high-grade gliomas is frequent. The molecular mechanisms are quite established, however the heterogeneity of glioblastomas force the scientist to look for the new therapeutic targets. The aim of the study was to evaluate the caspase-3 and survivin expression in correlation with MIB-1 expression in gliomas of various grade to assess the apoptosis in gliomas and to determinate new possible targets for the future therapy.
Material and methods:
We identified 131 patients with a histopathological diagnosis of astrocytic tumors (diffuse astrocytoma, anaplastic astrocytoma and glioblastoma). The evaluation of caspase-3, survivin and MIB-1 expression was done using immunohistochemical methods.
Results:
Caspase-3 and survivin expression was observed both in low- and high-grade astrocytomas. The differences in expression were the most evident in glioblastoma group. All primary glioblastomas (31 cases) expressed caspase-3. In secondary glioblastoma group only 17 out of 30 specimens were positive for caspase-3. Survivin expression was observed in 80.6% primary glioblastomas and in all examined secondary glioblastomas and the staining was strong and diffuse in all cases. MIB-1 expression was low in diffuse astrocytomas (DA) and ranged between 1 and 5%. In anaplastic astrocytoma group it was ranged between 5 and 10% and the highest percentage of the positive cells was observed in glioblastoma cases and ranged from 10% even to 30%. The most evident MIB-1 expression was observed in the cells surrounding the pathological blood vessels and necrosis.
Conclusions:
The high incidence of survivin and caspase-3 expression in diffuse and anaplastic astrocytoma cases may suggest, that the regulation between pro- and antiapoptotic proteins may play an important role in tumor growth and progression. The overexpression of survivin and MIB-1 expression in glioblastoma cases also may confirm the theory about the important role of anti-apoptotic and proliferation processes in glioblastoma progression and as such may be potential therapeutic targets.
Glioblastoma (GBM) is the most common and lethal brain tumor. Characterization of GBM with poor prognosis although new treatment modalities hasn't been changed up to date. New molecular targets in order to overcome resistance and recurrence are needed. Autophagy, a self-catabolic process, has two converse role in tumorigenesis, acting both as a tumor suppressor and a protector of cancer cell survival, tumor dormancy, progression, and therapeutic resistance. Autophagy with its dual role become target of cancer therapy. Midkine (MK), a growth factor with cytokine activity, is expressed highly during embryogenesis, but interestingly, MK is not detectable in healthy adults and only re-appears in the body as a part of the pathogenesis of diseases such as cancer. MK promotes proliferation, migration, anti-apoptotic manner, mitogenesis, transforming, and angiogenesis in various cells. In the central nervous system. High MK expression correlates with the progression of human astrocytomas, MK mRNA and protein expression levels were higher in high-grade astrocytomas as GBMs than in low-grade astrocytomas. MK correlates with the poor prognosis of GBM. This review is focused on the relationship between MK and autophagy in GBM and GBM treatment and pro-mising usage of MK in order to switch autophagy to cell death. © 2012 Springer Science+Business Media Dordrecht. All rights reserved.
Survivin, a member of the inhibitor of apoptosis (IAP) protein family that inhibits caspases and blocks cell death, is highly expressed in most cancers and is associated with a poor clinical outcome. Survivin has consistently been identified by molecular profiling analysis to be associated with high tumour grade cancers, different disease survival and recurrence. Polymorphisms in the survivin gene are emerging as powerful tools to study the biology of the disease and have the potential to be used in disease prognosis and diagnosis. The survivin gene polymorphisms have also been reported to influence tumour aggressiveness as well as survival of cancer patients. The differential expression of survivin in cancer cells compared to normal tissues and its role as a nodal protein in a number of cellular pathways make it a high target for different therapeutics. This review discusses the complex circuitry of survivin in human cancers and gene variants of survivin, and highlights novel therapy that targets this important protein.
Alternative splicing enables the generation of different proteins from a single gene, greatly increasing the use of genetic information. The resultant protein isoforms often have different biological properties effecting the phenotype of the cell in which it is expressed. Dysregulation of alternative splicing is a common occurrence in cancer and may lead to the formation of truncated or degraded proteins through the introduction of immature stop codons or nonsense mediated decay. Increasing evidence indicates that cancer-associated splicing variants play an important role in tumor initiation and progression. In this review, we summarize the evidence supporting the relevance of alternative splicing in glioblastoma multiforme (GBM). Specifically, we focus on the role of alternative splicing in GBM pathogenesis with an emphasis on the effect of aberrant alternative splicing of FGFR, GLI-1, and EGFR. The significance of exploiting alternatively spliced isoforms as potential biomarkers which may contribute to the development of diagnostic and prognostic methods, in addition to serving as molecular targets in GBM, will be discussed.
Survivin, an inhibitor of apoptosis protein, is a potentially prognostic factor and therapeutic target in breast carcinoma, but no consensus exists based on heterogeneous data. The aim of this present study is to clarify the prognostic relevance of survivin in breast cancer patients. Relevant articles were screened in PubMed and EMBASE databases. Patients’ clinical characteristics, overall survival (OS), disease/recurrence-free survival (DFS/RFS) and positive expressed survivin rates were extracted for further analysis. Statistics extracted from Kaplan–Meier survival curves were calculated indirectly with methods developed by Parmar, Williamson, and Tierney. Multivariate Cox hazard regression analysis data were used directly in Stata 11.0. Pooled hazard ratio (HR) and 95% confidence interval (CI) were calculated to evaluate the prognostic role of survivin in breast cancer. Online literature search identified 23 articles containing 3,259 breast cancer patients. Our meta-analysis of all included studies about survival outcomes showed positive correlation between poor prognosis and survivin expression. Pooled HRs (95% CIs) for OS and DFS/RFS were 1.37 (1.12–1.68) and 1.34 (1.02–1.76), respectively. Subgroup analyses considering methods used to detect survivin (immunohistochemistry or not) and localization of survivin (whole, nuclear or cytoplasm of the cell) were also conducted, and all the above analyses supported the stability of the prognostic role of survivin. In addition, our study revealed a significant association between survivin expression and lymph node metastasis (OR: 2.74; 95% CI: 1.27–5.93) or stage of breast cancer (OR: 2.01; 95% CI: 1.29–3.13). Positive expression of survivin demonstrated a significantly higher risk of recurrence and decreased OS rates in breast cancer.
Malignant gliomas (MGs) are among the most aggressive types of cancers in the human brain. Frequent tumor recurrence caused by a lack of effective therapeutic approaches results in a poor prognosis. Signal transducer and activator of transcription 3 (STAT3), an oncogenic protein, is constitutively activated in MGs and predicts a poor clinical outcome. STAT3 therefore is considered to be a promising target for the treatment of MGs. Cryptotanshinone (CTS), the main bioactive compound from the root of Salvia miltiorrhiza Bunge, has been reported to have various pharmacological effects. However, little is known about its function in MG cells. In this study, we evaluated the effect of CTS on the proliferation of human glioma cell lines (T98G and U87). Our results revealed that CTS significantly suppresses glioma cell proliferation. The phosphorylation of STAT3 Tyr705, but not Ser727, was inhibited by CTS, and STAT3 nuclear translocation was attenuated. Overexpression of constitutively active mutant STAT3C reversed the inhibitory effect of CTS, while knockdown STAT3 showed a similar inhibitory effect as CTS treatment. Following the downregulation of STAT3-regulated proteins cyclinD1 and survivin, cell cycle progression significantly arrested in G1/G0 phase. These results indicate that CTS may be a potential antiproliferation agent for the treatment of MGs and that its mechanism may be related to the inhibition of STAT3 signaling.
The aim of this study was to investigate the clinical significances of the mRNA expression of survivin gene and its four splice variants in the pathogenesis of colorectal cancer (CRC). CRC samples, matched adjacent tissues, and normal tissues were collected from surgical resections of 39 patients with histologically confirmed diagnosis. The mRNA expression of survivin and its four splice variants, that is, survivin-△Ex3, survivin-2B, survivin-3B, and survivin-2α, was detected using semiquantitative PCR and RT-PCR. Carcinoembryonic antigen (CEA) CAM5 was determined as control. The mRNA expression rates of survivin, survivin-△Ex3, survivin-2B, survivin-3B, surviving-2α, and CEA CAM5 in CRC samples were significantly higher than those in adjacent tissues (P < 0.01) and those in normal tissues (P < 0.01). The mRNA levels of the above variants in CRC samples were also significantly higher than those in adjacent tissues (P < 0.01) and those in normal tissues (P < 0.01). The mRNA levels of survivin, survivin-2B, and survivin-2α were not associated with any clinical variable of patients, while the levels of survivin-△Ex3 and survivin-3B were associated with lymphoid metastasis and Dukes grade (P < 0.05), and survivin-△Ex3 was associated with invasiveness. We concluded that mRNA expression rates and levels of survivin and its four splice variants elevated in CRC tissues, and expression levels of survivin-△Ex3 and survivin-3B were positively associated with tumor aggression.
Nuclear structure information available to August 1976 has been reviewed. The level properties obtained from various reaction and decay experiments are shown in the drawings. Experimental details, references, and comments are given in the text. The adopted level properties are summarized and referenced to systematics.
Survivin is a critical regulator of mitosis, and an inhibitor of apoptosis which is overexpressed in almost all cancers. In the current study, cell cycle profiles of normal proliferating human umbilical vein endothelial cells, prostate cancer, and lung cancer cell lines expressing varying levels of survivin and its splice variants were compared using a novel functional complementation assay. Defects in chromosome segregation and cytokinesis that were observed after depletion of endogenous survivin were not complemented by any of the survivin splice variants: survivin-2B, survivin-3B, survivin-ΔEx3, or survivin-2A when expressed exogenously at a level comparable to endogenous full-length survivin. Survivin variants were not detectable at the endogenous protein level. Cancer cells with higher levels of full-length survivin and survivin-2B expression, exhibited reduced caspase-3 activation following doxorubicin treatment and radiation. Whereas earlier studies focused on function and expression levels of survivin specific to cancer cells, the current study brings forward the essential role of survivin in normal dividing cells. Full-length survivin was found to be associated with Aurora-B kinase in the chromosomal passenger complex and depletion of survivin mimics mitotic phenotypes observed after Aurora-B kinase inhibition, in cancer as well as normal proliferating cells. Thus, our study establishes survivin as a marker of proliferation, rather than a cancer specific marker. Therefore, systemic therapeutic interventions targeting survivin will affect cancer as well as normal proliferating cells.
Alternative splicing of survivin (BIRC5) pre-mRNA generates the common isoform survivin and five additional splice variants: survivin2B, survivinΔEx3, survivin2α, survivin3B and survivin3α. Although the common isoform survivin has been shown to be involved in tumourigenesis and progression of various tumours, including diffusely infiltrating astrocytoma, the expression of other survivin splice variants in astrocytoma has not been fully investigated. Aims To evaluate the expression status and particularly prognostic significance of survivin splice variants in diffusely infiltrating astrocytoma.
mRNA expression of the six survivin variants in 73 diffusely infiltrating astrocytoma and 18 normal brain tissue samples was investigated by using reverse transcription-PCR (RT-PCR), and the total survivin protein expression by western analysis and immunohistochemistry. Association of survivin and its splice variants with tumour grade and prognosis was examined by statistical and survival analysis.
The survivin, survivinΔEx3, survivin2B and survivin2α splice variants were significantly elevated in diffusely infiltrating astrocytoma, and were barely detectable or not detectable at all in normal brain tissue. Survivin3B and survivin3α were not detected in these samples. The positive rates of the expressed four variants increased with astrocytoma grade. Significantly, expression of these splice variants was associated with much poorer disease-specific and progression-free survival (Kaplan-Meier analysis, p≤0.002). Cox regression model further indicated survivin2α mRNA expression and nuclear survivin immunostaining to be significant independent negative prognostic factors.
The survivin, survivinΔEx3, survivin2B and survivin2α splice variants were significantly elevated in astrocytoma, and were associated with tumour grade and poorer prognosis.
In previous cancer vaccine clinical trials targeting survivin, induction of specific CD8(+) T-cell responses did not consistently lead to clinical responses. Considering the critical role of CD4(+) T-cell help in generating antitumor immunity, integration of anti-survivin CD4(+) T-cell responses may enhance the efficacy of anti-survivin cancer immunotherapy. Human leukocyte antigen (HLA)-DP4 is emerging as an attractive MHC target allele of CD4(+) T cell-mediated immunotherapy, because it is one of the most frequent HLA alleles in many ethnic groups. In this article, we aimed to elucidate DP4-restricted CD4(+) T-cell responses against survivin in cancer patients.
We generated a human cell-based artificial antigen-presenting cell (aAPC) expressing HLA-DP4, CD80, and CD83 and induced DP4-restricted antigen-specific CD4(+) T cells. The number, phenotype, effector function, and in vitro longevity of generated CD4(+) T cells were determined.
We first determined previously unknown DP4-restricted CD4(+) T-cell epitopes derived from cytomegalovirus pp65, to which sustained Th1-biased recall responses were induced in vitro by using DP4-aAPC. In contrast, DP4-aAPC induced in vitro both Th1 and Th2 long-lived anti-survivin CD4(+) T cells from cancer patients. Both survivin-specific Th1 and Th2 cells were able to recognize survivin-expressing tumors in a DP4-restricted manner. Neither survivin-specific interleukin 10 secreting Tr1 cells nor Th17 cells were induced by DP4-aAPC.
DP4-restricted anti-survivin Th1 and Th2 immunity with sufficient functional avidity can be induced from cancer patients. The development of strategies to concurrently induce both CD4(+) and CD8(+) T-cell responses against survivin is warranted for optimal anti-survivin cancer immunotherapy.
To explore the immunogenicity of glioma stem-like cell-associated antigens (SAAs) from sorted or unsorted glioma tumor stem-like cells (TSCs) as well as irradiated TSCs. Two primary human malignant glioma lines (SHG62, SHG66) and U87 cell line were primarily cultured in the serum-free medium (SFM) supplemented with EGF/bFGF. TSCs were identified by their self-renewal, multi-lineage differentiation and tumorigenic activity. To prepare SAAs in vitro, CD133+ TSCs were sorted either by magnetic cell sorting or with irradiation (6 Gy).The cytotoxicity induced by autogenous myeloid dendritic cell (DC)-mediated SAA-specific cytotoxic T lymphocytes (CTLs) was assessed by the Just Another Method test. SHG62, SHG66, and U87 cells contained TSCs. CD133+ SAAs-specific CTLs were significantly more cytotoxic than effector cells loaded with unsorted SAA (P < 0.05). Effector cells loaded with irradiated SAAs were more cytotoxic than those with regular SAAs (P < 0.01). SAAs from CD133+ TSCs and irradiated TSCs provide highly immunogenic antigens. TSCs might be a novel source of antigens for DC vaccination against malignant gliomas.
Pelvic retroperitoneal liposarcomas are rare tumors of mesenchymal origin. We present a case of a giant pelvic well-differentiated liposarcoma of the retroperitoneum in a woman, presenting with a large abdominal mass.
A 62-year-old woman presented with a rapid abdominal enlargement. Pelvic examination revealed a huge mass occupying the whole pelvis and part of the right abdomen. At surgery, the pelvic organs were displaced to the right side by a retroperitoneal mass that marginally involved the part of the posterior abdominal wall, left parietal peritoneum of the diaphragm, left colic flexure and the left adnexa. The huge mass, uterus, and both adnexa were removed; hemicolectomy and latero-lateral anastomosis were carried out. No adjuvant irradiation was given. The patient is alive and free of disease five years after the operation.
The gynecologist should consider retroperitoneal liposarcoma in the differential diagnosis of large pelvic or abdominal masses.
Female genital graft-versus-host disease (GVHD) occurs after allogenic stem cell transplantation (SCT) in 25% of long-term survivors. To date, 28 severe cases with hematocolpos requiring surgery have been documented. We report two cases of severe gynecological GVHD. Although most of the vaginal GVH disease cases occurred within two years after oncologic treatment, our two cases highlight the possibility of a very long onset. We also confirmed that hormonal replacement therapy does not protect against synechia and that corticoid treatment has a good therapeutic value in recurrence after surgery. In conclusion, women with SCT experience a high risk of vaginal GVHD that could impair quality of life and sexual function. These cases pinpointed the importance of very long-term gynecological follow-up in order to diagnose early symptoms. To date, only early therapy with local corticoid application might reduce symptoms and development of severe genital GVHD. We support systematic use of topical corticoid therapy in severe cases even after surgery because of the high incidence of recurrence.
The aim of this study was to determine the role of survivin, caspase-3 and p53 expression in cervical cancer, and their correlation with clinicopathological features and prognosis.
Two hundred and twenty-eight cases of cervical disease were analyzed retrospectively between February 2003 and May 2007 at Taizhou Hospital of Taizhou Enze Medical Center. The expressions of survivin, caspase-3 and p53 were detected by immunohistochemistry (EnVision), assessed by tissue microarray. The correlation of the three genes and clinicopathological factors as well as prognosis were statistically analyzed.
The results showed that the positive expression rate of survivin, caspase-3 and p53 in cervical cancer was significantly higher than in the CIN group and cervicitis group (p < 0.05). The expression of survivin was related with clinical staging, stromal involvement and lymph node metastasis (p < 0.05). The positive ratio of caspase-3 was significantly different from histological grading (p < 0.05). The positive expression of p53 was correlated with histological type and grading (p < 0.05). The expression of survivin in cervical cancer was negatively associated with that of caspase-3 (p < 0.01). The positive expression of survivin in the survival group and non-survival group was significantly statistically difference (p < 0.01). There was a significant difference between survivin expression and survival duration by the log-rank method. Whereas no association with survival was seen for caspase-3 and p53 positivity.
Survivin, caspase-3 and p53 may play an important role in the occurrence and development of cervix carcinoma. It has been suggested that the high expression of survivin or p53 and low expression of caspase-3 are closely correlated with cervical cancer. They could be used as markers for malignant degree and invasiveness of cervix cancer. Survivin can also be used in the estimation of prognosis and survival time of cervix carcinoma.
The prognosis of patients with malignant glioma is poor in spite of multimodal treatment approaches consisting of neurosurgery, radiochemotherapy and maintenance chemotherapy. Among innovative treatment strategies like targeted therapy, antiangiogenesis and gene therapy approaches, immunotherapy emerges as a meaningful and feasible treatment approach for inducing long-term survival in at least a subpopulation of these patients. Setting up immunotherapy for an inherent immunosuppressive tumor located in an immune-privileged environment requires integration of a lot of scientific input and knowledge of both tumor immunology and neuro-oncology. The field of immunotherapy is moving into the direction of active specific immunotherapy using autologous dendritic cells (DCs) as vehicle for immunization. In the translational research program of the authors, the whole cascade from bench to bed to bench of active specific immunotherapy for malignant glioma is covered, including proof of principle experiments to demonstrate immunogenicity of patient-derived mature DCs loaded with autologous tumor lysate, preclinical in vivo experiments in a murine orthotopic glioma model, early phase I/II clinical trials for relapsing patients, a phase II trial for patients with newly diagnosed glioblastoma (GBM) for whom immunotherapy is integrated in the current multimodal treatment, and laboratory analyses of patient samples. The strategies and results of this program are discussed in the light of the internationally available scientific literature in this fast-moving field of basic science and translational clinical research.
Immunotherapy is a new light of hope for the treatment of malignant gliomas. The brain is no longer believed to be an immunologically privileged organ. The major advantage of immunotherapy is the tumor-specific cytotoxic effect on the tumor cells with minimal side effects. Autologous dendritic cells (DCs)-based immunotherapy is a promising and feasible method. DCs are the most potent antigen-presenting cells (APCs). DCs prime T lymphocytes by epitopic major histocompatibility (MHC) class I and II for CD8(+) cytotoxic T lymphocytes (CTLs) and CD4(+) T helper cells, respectively. From the tissue specimen examination after DCs-based immunotherapy, CD8(+) CTLs have replaced T regulatory cells (Tregs) as the major dominant tissue infiltrating lymphocytes (TILs). CD8(+) CTLs play a key role in the tumor response, which may also be effective against cancer stem cells. DCs themselves also produce many cytokines including interferon-gamma and interleukin (IL-2) to kill the tumor cells. From the preliminary better outcomes in the literature for malignant gliomas, DC-based immunotherapy may improve tumor response by increasing the survival rate and time. It is recommended that DC-based immunotherapy is applied as soon as possible with conjunctive radiotherapy and chemotherapy. Malignant gliomas have heterogeneity of tissue-associated antigens (TAAs). To find universal common antigens through different kinds of tumor culture may be the essential issue for tumor vaccine development in the future.
Inhibitors of programmed cell death (apoptosis) aberrantly prolonging cell viability may contribute to cancer by facilitating the insurgence of mutations and by promoting resistance to therapy. Despite the identification of several new apoptosis inhibitors related to bcl-2 or to the baculovirus IAP gene, it is not clear whether apoptosis inhibition plays a general role in neoplasia. Here, we describe a new human gene encoding a structurally unique IAP apoptosis inhibitor, designated survivin. Survivin contains a single baculovirus IAP repeat and lacks a carboxyl-terminal RING finger. Present during fetal development, survivin is undetectable in terminally differentiated adult tissues. However, survivin becomes prominently expressed in transformed cell lines and in all the most common human cancers of lung, colon, pancreas, prostate and breast, in vivo. Survivin is also found in approximately 50% of high-grade non-Hodgkin's lymphomas (centroblastic, immunoblastic), but not in low-grade lymphomas (lymphocytic). Recombinant expression of survivin counteracts apoptosis of B lymphocyte precursors deprived of interleukin 3 (IL-3). These findings suggest that apoptosis inhibition may be a general feature of neoplasia and identify survivin as a potential new target for apoptosis-based therapy in cancer and lymphoma.
Inhibitors of programmed cell death (apoptosis) may regulate tissue differentiation and aberrantly promote cell survival in neoplasia. A novel apoptosis inhibitor of the IAP gene family, designated survivin, was recently found in all of the most common human cancers but not in normal, terminally differentiated adult tissues. The expression of survivin in embryonic and fetal development was investigated. Immunohistochemistry and in situ hybridization studies demonstrated strong expression of survivin in several apoptosis-regulated fetal tissues, including the stem cell layer of stratified epithelia, endocrine pancreas, and thymic medulla, with a pattern that did not overlap with that of another apoptosis inhibitor, bcl-2. A sequence-specific antibody to survivin immunoblotted a single approximately 16.5-kd survivin band in human fetal lung, liver, heart, kidney, and gastrointestinal tract. In mouse embryo, prominent and nearly ubiquitous distribution of survivin was found at embryonic day (E)11.5, whereas at E15 to -21, survivin expression was restricted to the distal bronchiolar epithelium of the lung and neural-crest-derived cells, including dorsal root ganglion neurons, hypophysis, and the choroid plexus. These data suggest that expression of survivin in embryonic and fetal development may contribute to tissue homeostasis and differentiation independently of bcl-2. Aberrations of this developmental pathway may result in prominent re-expression of survivin in neoplasia and abnormally prolonged cell viability.
A novel inhibitor of apoptosis designated survivin has recently been found in many common human cancers but not in normal tissues. A potential distribution of survivin in gastric cancer and its implication for apoptosis inhibition have been investigated. Recombinant survivin expressed in Escherichia coli as a glutathione S-transferase fusion protein was used to raise a novel panel of mouse monoclonal antibodies. In an immunohistochemical analysis of 174 cases of gastric carcinomas (stages I-III), anti-survivin monoclonal antibody 8E2 (IgG1) reacted with 34.5% of cases (60 of 174 cases) with a variable number of tumor cells stained (20-100%). In contrast, no expression of survivin in neighboring normal tissues was observed. When stratified for p53 and bcl-2 expression and apoptotic index, the expression of survivin significantly segregated with p53- and bcl-2-positive cases [56.1 versus 15.2% (P = 0.001) and 69.2 versus 31.6% (P = 0.006), respectively] and with a decreased apoptotic index as compared with that of survivin-negative tumors (0.97 +/- 0.64 versus 0.62 +/- 0.39%, P < 0.001). These data identify a role for survivin in promoting aberrantly increased cell viability in gastric cancer and suggest a potential correlation between accumulated p53 and survivin expression in neoplasia.
Progression of the cell cycle and control of apoptosis (programmed cell death) are thought to be intimately linked processes, acting to preserve homeostasis and developmental morphogenesis. Although proteins that regulate apoptosis have been implicated in restraining cell-cycle entry and controlling ploidy (chromosome number), the effector molecules at the interface between cell proliferation and cell survival have remained elusive. Here we show that a new inhibitor of apoptosis (IAP) protein, survivin, is expressed in the G2/M phase of the cell cycle in a cycle-regulated manner. At the beginning of mitosis, survivin associates with microtubules of the mitotic spindle in a specific and saturable reaction that is regulated by microtubule dynamics. Disruption of survivin-microtubule interactions results in loss of survivin's anti-apoptosis function and increased caspase-3 activity, a mechanism involved in cell death, during mitosis. These results indicate that survivin may counteract a default induction of apoptosis in G2/M phase. The overexpression of survivin in cancer may overcome this apoptotic checkpoint and favour aberrant progression of transformed cells through mitosis.
The inhibitor of apoptosis protein family has been characterized over the past 5 years, initially in baculovirus and more recently in metazoans. The IAPs are a widely expressed gene family of apoptotic inhibitors from both phylogenic and physiologic points of view. The diversity of triggers against which the IAPs suppress apoptosis is greater than that observed for any other family of apoptotic inhibitors including the bcl-2 family. The central mechanisms of IAP apoptotic suppression appear to be through direct caspase and pro-caspase inhibition (primarily caspase 3 and 7) and modulation of and by the transcription factor NF-kappaB. Although evidence for a direct oncogenic role for the IAPs has yet to be delineated, a number of lines of evidence point towards this class of protein playing a role in oncogenesis. The strongest evidence for IAP involvement in cancer is seen in the IAP called survivin. Although not observed in adult differentiated tissue, survivin is present in most transformed cell lines and cancers tested to date. Survivin has been shown to inhibit caspase directly and apoptosis in general, moreover survivin protein levels correlate inversely with 5 year survival rates in colorectal cancer. Recent data has also implicated survivin in cell cycle control. The second line of evidence for IAP involvement in cancer comes from their emerging role as mediators and regulators of the anti-apoptotic activity of v-Rel and NF-kappaB transcription factor families. The IAPs have been shown to be induced by NF-kappaB or v-Rel in multiple cell lines and conversely, HIAP1 and HIAP2 have been shown to activate NF-kappaB possibly forming a positive feed-back loop. Overall a picture consistent with an IAP role in tumour progression rather than tumour initiation is emerging making the IAPs an attractive therapeutic target.
Recently, a novel antiapoptosis gene, i.e., survivin, was identified as a structurally unique member of the inhibitor of apoptosis protein family. Survivin expression is turned off during fetal development and not found in non-neoplastic adult human tissues but is again turned on in the most common human cancers. The antiapoptotic properties of survivin might provide a significant growth advantage in tumors and possibly also contribute to chemoresistance of cancer. Therefore, we analyzed the expression of survivin in human renal cell carcinomas (RCCs), known to be largely resistant to chemotherapy. Northern blot analysis and RT-PCR revealed survivin expression in newly established RCC cell lines (n = 11) of all major histological types. Moreover, we identified two novel splice variants of survivin, lacking exon 3 (survivin-deltaEx3) or retaining a part of intron 2 as a cryptic exon (survivin-2B). Both sequence alterations cause marked changes in the structure of the corresponding proteins, including structural modifications of the baculovirus inhibitor of apoptosis protein repeat domain. The role of the novel isoforms in the regulation of apoptosis was assessed in transfection experiments, showing conservation of antiapoptotic properties for survivin-deltaEx3 and a markedly reduced antiapoptotic potential for survivin-2B. In conclusion, our observations suggest a complex regulatory balance between the different isoforms of survivin, which might determine the response to proapoptotic stimuli, not only in human RCCs but also in fetal tissues and other types of cancer.
Aberrant inhibition of programmed cell death (apoptosis) prevents normal homeostasis and promotes tissue tumorigenesis, but whether it also influences the outcome of common cancers has remained arguable. The expression of a novel IAP apoptosis inhibitor, survivin, in breast cancer and its association with tumor cell apoptosis and overall prognosis were examined in this study. Immunohistochemical analysis showed that survivin expression was positive in 118 of 167 cases (70.7%) of breast carcinomas of histological stages I to IH. In contrast, no expression of survivin in adjacent normal tissue was detected. Although survivin expression was not correlated with p53 mutations, survivin-positive cases were strongly associated with bcl-2 expression (78.0% versus 47.5%; P = 0.0005) and reduced apoptotic index (0.62% +/- 0.51% versus 1.27% +/- 1.37%; P < 0.0001). In addition, patients with low apoptotic index (<0.52%) had worse survival rates than the group with high apoptotic index (> or =0.52%; P = 0.028), and multivariate Cox proportional hazard model analysis identified apoptotic index as an independent prognostic factor (P = 0.024). The results suggest that apoptosis inhibition by survivin, alone or in cooperation with bcl-2, is a significant prognostic parameter of worse outcome in breast carcinoma.
Mechanisms controlling endothelial cell survival during angiogenesis were investigated. Stimulation of quiescent endothelial cells with mitogens, including vascular endothelial growth factor and basic fibroblast growth factor, induced up to approximately 16-fold up-regulation of the cell cycle-regulated apoptosis inhibitor survivin. Mitogen stimulation rapidly increased survivin RNA expression in endothelial cells, which peaked after 6 to 10 hours in culture and decreased by 24 hours. Inflammatory cytokines, tumor necrosis factor alpha, and interleukin-1 did not induce survivin expression in endothelial cells. Formation of three-dimensional vascular tubes in vitro was associated with strong induction of survivin in endothelial cells, as compared with two-dimensional cultures. By immunohistochemistry, survivin was minimally expressed in endothelium of nonproliferating capillaries of normal skin, whereas it became massively up-regulated in newly formed blood vessels of granulation tissue in vivo. Recombinant expression of green fluorescent protein survivin in endothelial cells reduced caspase-3 activity and counteracted apoptosis induced by tumor necrosis factor alpha/cycloheximide. These findings identify survivin as a novel growth factor-inducible protective gene expressed by endothelial cells during angiogenesis. Therapeutic manipulation of survivin expression and function in endothelium may influence compensatory or pathological (tumor) angiogenesis.
Survivin (SVV) is a family member of inhibitor of apoptosis proteins (IAPs) and its expression is cell cycle regulated. The gene is mapped to chromosome 17q25, the region of which is frequently gained in advanced stages of neuroblastoma (NBL). However, the role of SVV in NBL is poorly understood. Here we studied the clinical and biological role of SVV in NBL. A 1.9 kb SVV transcript was expressed in all of 9 NBL cell lines at higher levels than those in adult cancer cell lines. In 34 primary NBLs, high levels of SVV expression was significantly associated with age greater than 12 months (two sample t-test: P= 0.0003), advanced stages (P = 0.0136), sporadic tumors (P= 0.0027) and low levels of TrkA expression (P = 0.0030). In NBL cell lines, SVV mRNA expression was dramatically down-regulated in CHP134 and IMR32 cells undergoing apoptosis after treatment with all-trans retinoic acid (RA) or serum deprivation. It was only moderately decreased in cells (SH-SY5Y and CHP901) undergoing RA-induced differentiation. On the other hand, in proliferating NBL cells or RA-treated SK-N-AS line which is refractory to RA, the SVV mRNA remained at steady state levels or rather up-regulated. Furthermore, transfection of SVV into CHP134 cells induced remarkable inhibition of the RA-induced apoptosis. Collectively, our results suggest that high expression of SVV is a strong prognostic indicator for the advanced stage neuroblastomas, and that it could be one of the candidate genes for the 17q gain.
Survivin is a novel inhibitor of apoptosis. It has been reported that survivin is expressed during fetal development and in cancer tissues, but its expression has not been reported in adult tissues. We investigated the expression of survivin in the endometria of women with regular menstrual cycles using reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry, and compared these findings with Bcl-2, an apoptosis inhibitor. Survivin mRNA was detected by RT-PCR in all samples (nine of nine) of endometrium during the secretory phase, but in only four out of seven samples from endometrium during the proliferative phase, and in none of the atrophic endometrium. Immunohistochemistry demonstrated a survivin protein expression that was strongest in the nuclei of glandular epithelial cells during the late secretory phase. In the proliferative phase, glandular epithelial cells were not stained for survivin. The cyclic changes of survivin and Bcl-2 showed an inverse relationship, with Bcl-2 expression being strongest in the proliferative phase and survivin expression being strongest in the secretory phase. The up-regulation of survivin expression may be due to the concurrent rise in progesterone concentrations during the normal menstrual cycle. Moreover, survivin could play an important role independent of Bcl-2 in physiological homeostasis in the normal endometrium.
Procedure
We investigated the expression of survivin (SVV) and its isoform (SVV‐β/2B) during different biological properties in neuroblastoma (NBL).
Results
High levels of SVV mRNA expression were significantly associated with advanced stages of NBL, diagnosis at over 1 year of age, low levels of TrkA expression, and sporadic tumors. Expression of a novel isoform, SVV‐β/2B, which had an insertion of 23 amino acids within the unique BIR domain was predominant in some favorable NBLs, while it was low and ubiquitous in most normal and malignant tissues. The SVV expression wasdown‐regulated during apoptosis induced by retinoic acid (RA) in CHP134 NBL cells, which was inhibited by forced expression of SVV. In contrast, SVV‐β was constantly expressed during apoptosis. Like SVV ,SVV‐β was also highly expressed during G2/M in a cell cycle‐dependent manner, and was associated with but competed against SVV for binding with polymerized tubulin.
Conclusion
These data suggest that expression of SVV is a poor prognostic indicator in human NBL, and it promotes growth and survival by regulating the levels of both isoforms. Med. Pediatr. Oncol. 35:550–553, 2000. © 2000 Wiley‐Liss, Inc.
BACKGROUND/AIMS
Inhibition of programmed cell death (apoptosis) is associated with increased tumour aggressiveness, and expression ofSurvivin, an antiapoptosis gene, in colorectal carcinomas may provide important prognostic information.
PATIENTS/METHODS
Expression of Survivin messenger RNA was evaluated by reverse transcription-polymerase chain reaction in 144 colorectal carcinomas and 86 adjacent histologically normal mucosa samples from patients for whom long term follow up data were available.
RESULTS
Survivintranscripts were detected in a significantly greater proportion of carcinomas (63.5%) than normal mucosa samples (29.1%; p<0.001). The prevalence of Survivin expression was independent of advancing pathological stage. Death due to recurrent cancer following curative resection was predicted independently by tumour expression of Survivin (hazard ratio (HR) 2.60; 95% confidence interval (95% CI) 1.17–5.75) and lymph node metastases (HR 2.38; 95% CI 1.21–4.70). On stage wise analysis, the predictive value of Survivin expression was limited to patients with stage II colorectal carcinomas; those with Survivin negative tumours had a five year survival rate of 94.4% compared with 44.8% for patients withSurvivin positive tumours (p=0.004, log rank test).
CONCLUSION
In patients with stage II colorectal carcinomas,Survivin expression provides prognostic information that may have important therapeutic implications.
Survivin, a recently identified inhibitor of apoptosis protein (IAP), is expressed in diverse embryonic tissues and in various human cancers. We have investigated the quantitative expression of survivin mRNA by a sensitive TaqMan™-based RT-PCR assay in tissue samples from 94 patients with soft tissue sarcomas (STS). Survivin transcript levels were measured and normalized to GAPDH transcripts. By using a multivariate Cox regression analysis, we found an inverse correlation between the level of survivin mRNA (ratio >2 zmol survivin/amol GAPDH) and the rate of overall survival (p = 0.009, RR = 2.7). Survivin transcript variants as detected by qualitative RT-PCR analysis were revealed in 36 of 56 STS patients (64%). Only survivin ΔEx3 and/or full-length survivin variants but not survivin 2B were identified. Our results suggest that a higher level of survivin mRNA is an independent predictor of survival for STS patients. © 2001 Wiley-Liss, Inc.
Regulation of the homeostatic balance between cell proliferation and cell death is essential for development and maintenance of multicellular organisms. Physiologic, or programmed, cell death is dependent on a genetically encoded and evolutionarily conserved pathway that induces a form of cellular suicide known as apoptosis. In the past decade, it has become clear that the regulatory mechanisms controlling programmed cell death are as fundamental, and as complex, as those regulating cell proliferation. Perturbation of the signaling cascades regulating apoptosis, whether by extracellular triggers, acquired or germline genetic mutations, or viral mimicry of signaling molecules, can result in a wide variety of human diseases. Analysis of these regulatory pathways has led to a better understanding of the etiology and pathogenesis of many human diseases, notably cancers, infectious diseases including AIDS, autoimmune diseases, and neurodegenerative/neurodevelopmental diseases. Our understanding of the regulation of programmed cell death in health and disease is far from complete, and the challenge of converting that understanding into new therapeutic modalities has only begun to be approached.
Deregulated inhibition of apoptosis (programmed cell death) may facilitate the insurgence of neoplasia, but whether it also influences the outcome of common cancers has remained controversial. In this study, we investigated the expression of a novel inhibitor of apoptosis, survivin, in colorectal cancer and its relationship with tumor cell apoptosis and overall prognosis. By immunohistochemistry, survivin was expressed in 91 of 171 (53.2%) cases of colorectal carcinomas of histological stages 0 to IV. In contrast, normal colon epithelium did not express survivin. Although survivin expression did not correlate with p53 abnormalities (46.5% versus 58.0%; P = 0.18), survivin-positive cases were strongly associated with bcl-2 expression (72.5% versus 27.4%; P < 0.0001) and reduced apoptotic index (0.76% +/- 0.39% versus 1.17% +/- 0.62%; P < 0.0001). Expression of survivin alone in bcl-2-negative (discordant) cases also resulted in reduced apoptotic index (0.82% +/- 0.57% versus 1.16% +/- 0.66%; P = 0.0046). When analyzed for prognostic significance, patients with low apoptotic index (< 0.97%) had worse survival rates than the group with high apoptosis (P < 0.001), and a multivariate Cox proportional hazard model identified reduced apoptosis as an independent predictive factor for overall survival (P < 0.0001). These data demonstrate that apoptosis inhibition by survivin, alone or in cooperation with bcl-2, is an important predictive/prognostic parameter of poor outcome in colorectal carcinoma and identify survivin as a new diagnostic/therapeutic target in cancer.
Apoptosis plays a fundamental part in epidermal homeostasis, and apoptotic cells have been detected in normal and diseased skin. Little is known, however, on the inhibitory mechanisms of apoptosis at the skin level. In addition to bcl-2, a novel inhibitor of apoptosis designated survivin and structurally analogous to IAP apoptosis inhibitors has been recently identified. The expression of survivin in normal and pathologic skin was investigated. Immunohistochemical studies revealed that survivin is expressed in basal keratinocytes, but not in suprabasal epidermal layers, with a pattern similar to bcl-2. In western blots, the anti-survivin antibody recognized a single band of 16.5 kDa in protein extracts from normal human keratinocytes in culture, in agreement with the predicted size of survivin. In addition, survivin immunoreactivity was detected in benign and malignant melanocytic lesions, with strong expression in invasive lesions of melanomas. Whereas survivin staining was undetectable in benign epithelial tumors, such as seborrheic keratoses, it was observed in all epidermal layers in Bowen's disease. Interestingly, at variance with bcl-2, survivin was markedly expressed in squamous cell carcinoma, but virtually lacking in basal cell carcinoma, suggesting that these two apoptosis inhibitors may act through different anti-apoptotic pathways. Deregulation of survivin may influence both epidermal homeostasis and the development of melanoma and nonmelanoma skin cancer.
The survivin gene is a novel apoptosis inhibitor, related to the baculovirus gene, which is believed to play a pivotal role in fetal development and in cancer. We hypothesised that survivin would be expressed in tumors of patients with non-small-cell lung cancer (NSCLC), and we attempted to determine the influence of survivin re-expression on clinical outcome in patients with up to stage IIIA NSCLC who had undergone radical surgery.
We designed a reverse transcriptase polymerase chain reaction (RT-PCR) assay to study the expression of the survivin gene in 83 NSCLC tumor samples and compared the results with relevant clinical and pathologic data.
The RT-PCR identified survivin gene transcript in 71 (85. 5%) of the tumor samples and in only 10 (12%) of the paired, histopathologically normal lung samples. There was no relationship between histologic subtype (squamous v nonsquamous) and survivin gene expression. The 12 patients without survivin expression had significantly better overall survival than the 71 patients with survivin expression (P =.01 by univariate analysis; relative risk, 2. 1). There was no significant correlation between survivin expression and age, sex, cigarette smoking, histologic subtype, tumor differentiation, tumor size, or the presence of mediastinal lymph node metastases in surgical specimens.
The survivin gene was expressed in a vast majority of NSCLC tumors. We conclude that survivin transcript is a defining diagnostic marker for NSCLC that may also yield prognostic information and, as an apoptosis inhibitor, be an important target in cancer therapy.
Inhibition of programmed cell death (apoptosis) is associated with increased tumour aggressiveness, and expression of Survivin, an antiapoptosis gene, in colorectal carcinomas may provide important prognostic information.
Expression of Survivin messenger RNA was evaluated by reverse transcription-polymerase chain reaction in 144 colorectal carcinomas and 86 adjacent histologically normal mucosa samples from patients for whom long term follow up data were available.
Survivin transcripts were detected in a significantly greater proportion of carcinomas (63.5%) than normal mucosa samples (29.1%; p<0.001). The prevalence of Survivin expression was independent of advancing pathological stage. Death due to recurrent cancer following curative resection was predicted independently by tumour expression of Survivin (hazard ratio (HR) 2.60; 95% confidence interval (95% CI) 1. 17-5.75) and lymph node metastases (HR 2.38; 95% CI 1.21-4.70). On stage wise analysis, the predictive value of Survivin expression was limited to patients with stage II colorectal carcinomas; those with Survivin negative tumours had a five year survival rate of 94.4% compared with 44.8% for patients with Survivin positive tumours (p=0. 004, log rank test).
In patients with stage II colorectal carcinomas, Survivin expression provides prognostic information that may have important therapeutic implications.
Survivin is a recently described inhibitor of apoptosis. Because suppression of apoptosis is important for carcinogenesis and tumor growth, we investigated the expression and function of survivin in human hepatocellular carcinomas (HCCs). We have shown that 4 HCC cell lines and 7 out of 8 human HCC tissues expressed survivin messenger RNA (mRNA), whereas expression of survivin mRNA was not detected in normal liver and nontumor areas of these tissues using the reverse transcription polymerase chain reaction. Survivin was detected primarily in the nucleus by immunofluorescence staining of HCC cells. In addition, 14 of 20 (70%) HCC tissues showed positive nuclear staining for survivin, whereas nontumor tissues showed little detectable staining by immunohistochemistry. Survivin expression strongly correlated with the proliferation index but not significantly with the apoptosis index in HCC tissues. Therefore, we performed cell cycle analysis after survivin transfection and showed that overexpression of survivin resulted in a decrease in the G(0)/G(1) phase and an increase in the S phase in all 4 HCC cell lines. Furthermore, we have found that survivin interacted with cyclin-dependent kinase 4 (Cdk4) and overexpression of survivin released p21(WAF1/Cip1) (p21) from Cdk4. From these results, we conclude that survivin promotes cell proliferation by interacting with Cdk4 and releasing p21 from Cdk4. This may play an important role in carcinogenesis and progression of human HCCs.
Survivin is a newly discovered inhibitor of the apoptosis protein, IAP, expressed during development and in human cancers. The effector cell protease receptor-1 (EPR-1) gene is oriented in the opposite direction on the same DNA double strand. Thus, the Survivin and EPR-1 (Survivin/EPR-1) genes exist in a head-to-head configuration. It is not clear whether mutual expression of the Survivin/EPR-1 genes occurs in both normal cells and cancer cells. Here, we investigated the mutual expression of the Survivin/EPR-1 genes in 12 normal peripheral blood (PB) specimens, seven normal bone marrow (BM) specimens, five lymph node (LN) specimens, and seven leukemic cell lines, and 27 patients with malignant lymphoma (ML), four with acute lymphocytic leukemia (ALL), three with acute myelocytic leukemia (AML), and four with chronic myelocytic leukemia in blastic crisis (CML-BC). Using Northern blot analysis, small amounts of EPR-1 mRNA were detected in normal PB, normal BM and LN specimens, but no Survivin mRNA was detected. However, Survivin mRNA was detected in two of the 12 normal PB, six of the seven normal BM and one of the five LN specimens using reverse transcription and polymerase chain reaction (RT-PCR). Expression of both the Survivin and EPR-1 genes was detected in six of the seven cell line samples by Northern blot, and in all of them by RT-PCR. Mutual expression of the Survivin and EPR-1 genes was detected in three of the four CML-BC samples, 15 of the 27 ML samples, two of the four ALL samples, and all three AML samples using the RT-PCR method. No EPR-1 expression with or without Survivin expression was clearly detected in eight of the nine diffuse large B-cell lymphoma (DLB) specimens, two of the six follicular center lymphoma specimens, one of the four specimens of nodular sclerosis of Hodgkin's lymphoma, two of the four ALL specimens or one of the four CML-BC specimens. The data presented here show that disrupted expression of the Survivin/EPR-1 genes occurred in many kinds of hematologically malignant cells. This may be of biological importance.
We examined whether survivin acts as a constitutive and inducible radioresistance factor in pancreatic cancer cells. Using a quantitative TaqMan reverse transcription-polymerase chain reaction for survivin mRNA in five pancreatic cancer cell lines, we found an inverse relationship between survivin mRNA expression and radiosensitivity. PANC-1 cells, which had the highest survivin mRNA levels, were most resistant to X-irradiation; MIAPaCa-2 cells, which showed the least survivin mRNA expression, were the most sensitive to X-irradiation. Our results suggested that survivin could act as a constitutive radioresistance factor in pancreatic cancer cells. To determine whether radioresistance is enhanced by induction of survivin expression by irradiation, PANC-1 and MIAPaCa-2 cells were subjected to sublethal doses of X-irradiation followed by a lethal dose. Survivin mRNA expression was increased significantly in both PANC-1 and MIAPaCa-2 cell lines by pretreatment with a sublethal dose of X-irradiation, as was cell survival after exposure to the lethal dose. In this system, enzymatic caspase-3 activity was significantly suppressed in cells with acquired resistance. These results suggest that survivin also acts as an inducible radioresistance factor in pancreatic cancer cells. Survivin, then, appears to enhance radioresistance in pancreatic cancer cells; inhibition of survivin mRNA expression may improve the effectiveness of radiotherapy.
We investigated the expression of survivin (SVV) and its isoform (SVV-beta/2B) during different biological properties in neuroblastoma (NBL).
High levels of SVV mRNA expression were significantly associated with advanced stages of NBL, diagnosis at over 1 year of age, low levels of TrkA expression, and sporadic tumors. Expression of a novel isoform, SVV-beta/2B, which had an insertion of 23 amino acids within the unique BIR domain was predominant in some favorable NBLs, while it was low and ubiquitous in most normal and malignant tissues. The SVV expression wasdown-regulated during apoptosis induced by retinoic acid (RA) in CHP134 NBL cells, which was inhibited by forced expression of SVV. In contrast, SVV-beta was constantly expressed during apoptosis. Like SVV,SVV-beta was also highly expressed during G(2)/M in a cell cycle-dependent manner, and was associated with but competed against SVV for binding with polymerized tubulin.
These data suggest that expression of SVV is a poor prognostic indicator in human NBL, and it promotes growth and survival by regulating the levels of both isoforms.
A role of apoptosis (programmed cell death) in tumor formation and growth was investigated by targeting the apoptosis inhibitor survivin in vivo. Expression of a phosphorylation-defective survivin mutant (Thr(34)-->Ala) triggered apoptosis in several human melanoma cell lines and enhanced cell death induced by the chemotherapeutic drug cisplatin in vitro. Conditional expression of survivin Thr(34)-->Ala in YUSAC2 melanoma cells prevented tumor formation upon s.c. injection into CB.17 severe combined immunodeficient-beige mice. When induced in established melanoma tumors, survivin Thr(34)-->Ala inhibited tumor growth by 60-70% and caused increased apoptosis and reduced proliferation of melanoma cells in vivo. Manipulation of the antiapoptotic pathway maintained by survivin may be beneficial for cancer therapy.
In this study we investigated expression profiles of the anti-apoptotic gene survivin in malignant human haematopoietic cells.
Using a quantitative TaqMan reverse transcription-polymerase chain reaction, survivin and bcl-2 mRNA expression were examined in 12 malignant haematopoietic cell lines, in 21 patients with haematopoietic malignancies and in normal leukocyte fractions.
Survivin mRNA levels, demonstrable in all 12 malignant cell lines, differed but showed no relationship to the cell of origin. Conversely, no survivin mRNA expression was detected in normal leukocyte fractions. Further, survivin mRNA was expressed in 16 out of 21 patients with malignancies. Five days after treatment of HL-60 cells with a combination of all-trans retinoic acid and tumor necrosis factor, survivin expression decreased to 14.1% of that in untreated cells. Further, survivin mRNA expression in K-562/ADR cells with acquired resistance to adriamycin was 1.7 times that in parent K-562 cells.
Our results indicated the possibility that quantitation of survivin mRNA expression is a useful tool for the detection of haematopoietic tumor cells in clinical laboratory test and that survivin could be a target for treatment of haematopoietic malignancies.
Using homology searches, we identified a novel human inhibitor of apoptosis (IAP) gene. This gene has two splicing variants that contain open reading frames of 298 and 280 amino acids and both contained a single copy of baculovirus IAP repeat (BIR) and RING domain. We refer here to the longer and shorter variants as Livin alpha and beta, respectively. Semiquantitative reverse transcriptase-polymerase chain reaction demonstrated a tissue-specific and non-correlated expression pattern in both adult and fetal tissues. Both mRNA variants were detected in various transformed cell lines. Despite their very close similarity, the two isoforms have different antiapoptotic properties. Both isoforms have a significant antiapoptotic activity in the Jurkat T cell line after triggering apoptosis via tumor necrosis factor and CD95 receptors. The Livin alpha but not beta protects cells from apoptosis induced by staurosporine, but in contrast, apoptosis initiated by etoposide was blocked only by the beta isoform. This difference in biological activities may indicate the presence of critical amino acids outside the BIR and RING domains. These functional and tissue distribution differences of Livin alpha and beta suggest that Livin may play a complex role in the regulation of apoptosis.
Survivin, a recently identified inhibitor of apoptosis protein (IAP), is expressed in diverse embryonic tissues and in various human cancers. We have investigated the quantitative expression of survivin mRNA by a sensitive TaqMan-based RT-PCR assay in tissue samples from 94 patients with soft tissue sarcomas (STS). Survivin transcript levels were measured and normalized to GAPDH transcripts. By using a multivariate Cox regression analysis, we found an inverse correlation between the level of survivin mRNA (ratio >2 zmol survivin/amol GAPDH) and the rate of overall survival (p = 0.009, RR = 2.7). Survivin transcript variants as detected by qualitative RT-PCR analysis were revealed in 36 of 56 STS patients (64%). Only survivin DeltaEx3 and/or full-length survivin variants but not survivin 2B were identified. Our results suggest that a higher level of survivin mRNA is an independent predictor of survival for STS patients.
Survivin is a novel antiapoptotic gene that has been recently cloned and characterized. Its expression has been found to be of prognostic significance in several tumor types. This is the first study on the prognostic significance of survivin expression in human gliomas.
We used quantitative Western blot analysis with densitometry to determine survivin protein expression levels in 92 glioma cases for which frozen tissue was available for analysis. Survivin positivity and expression levels were correlated with histopathologic features of the tumors, apoptosis (as measured by cleaved, or activated, caspase 3 levels), and clinical outcome.
Survivin expression has clear prognostic value in human gliomas. Patients with detectable survivin expression had significantly shorter overall survival times (P <.0001) compared with those without detectable expression when all glioma patients were considered. Although glioblastoma multiforme (GBM) patients had significantly higher rates of survivin positivity and higher levels of survivin expression (P <.0001) than their non-GBM counterparts, the prognostic value of survivin expression seemed to be independent of histology alone. Survivin-positive GBM patients had significantly shorter overall survival times compared with survivin-negative GBM patients (P <.0001). Likewise, survivin-positive non-GBM patients had shorter survival times compared with survivin-negative non-GBM patients (P =.029). Furthermore, increasing levels of survivin expression significantly correlated with reduced survival times when all glioma patients were considered, and markedly so for GBM patients (P <.0001). Increasing survivin levels significantly correlated with reduced expression of cleaved caspase 3, indicating its association with antiapoptotic activity.
Survivin positivity and protein expression levels, as determined quantitatively, are of significant prognostic value in human gliomas and seem to be associated with reduced apoptotic capacity of these tumors.
To date an increasing number of T-cell epitopes derived from various tumor-associated antigens have been reported, and they proved to play significant roles for tumor rejection both in vivo and in vitro. Survivin was originally identified as a member of the inhibitor of apoptosis protein family. Expression of this gene is developmentally regulated. Although survivin is expressed during normal fetal development, the expression is barely detected in terminally differentiated adult tissues except for testis, thymus, and placenta. In contrast, it is abundantly expressed in a wide variety of malignant tissues. We examined the expression of survivin and the two splicing variants survivin-2B and survivin-DeltaEx3 in various cancer cells, immortalized cells, and normal adult tissues. It was demonstrated that two splicing variants were detected in various types of cancer cells as well as survivin, and their expression was more restricted to cancer cells as compared with survivin expression. To identify HLA-A24-restricted T-cell epitopes from survivin and the variant proteins, three peptides were selected from amino acid sequence of these proteins, based on the HLA-A24-binding motif. Peptide binding assay to HLA-A24 revealed that only one peptide designated as survivin-2B80-88 (AYACNTSTL) was capable of binding to HLA-A24. By stimulating peripheral blood lymphocytes with the peptide-pulsed antigen-presenting cells, CTLs were successfully induced in vitro from five of five HLA-A24-positive cancer patients. The CTLs showed significant cytotoxicity against HLA-A24-positive survivin-2B-positive cancer cells. These data suggest that survivin-2B80-88 may be a potent T-cell epitope eliciting CTL response against a splicing variant survivin-2B, which is specifically expressed in many kinds of cancer cells.
Survivin, a novel member of the inhibitor of apoptosis protein (IAP) family, reduces the susceptibility of tumor cells to proapoptotic stimuli, thereby promoting tumor cell survival during tumor progression and treatment with anticancer drugs. Recently, we identified 2 novel alternative splice variants of survivin, survivin-2B and survivin-Delta Ex3, which differ in their antiapoptotic properties. Survivin-2B has lost its antiapoptotic potential and may act as a naturally occurring antagonist of antiapoptotic survivin and survivin-Delta Ex3. Because the in vivo expression of these splice variants in human cancer has not been analyzed so far, 57 renal cell carcinomas (RCCs) were explored using quantitative reverse transcriptase polymerase chain reaction. We found that all RCCs express survivin-Delta Ex3, survivin-2B and survivin, the latter being the dominant transcript. When we compared early and intermediate stages with late stages of clear cell RCCs, no significant changes in the expression levels of survivin and survivin-Delta Ex3 became evident. However, a significant decrease was observed for the mRNA ratio between survivin-2B and survivin in late tumor stages (p = 0.036). Chromophilic/papillary RCCs, which are known to be less aggressive than clear cell RCCs, did not show significantly lower expression levels of antiapoptotic survivin and survivin-Delta Ex3, compared with stage-adjusted clear cell RCCs. Our study demonstrates for the first time in vivo expression of functionally different survivin variants and suggests a role of these survivin splice variants in the progression and clinical behavior of human RCCs.
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