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Quantitatively Determined Survivin Expression Levels Are of Prognostic Value in Human Gliomas
Abstract
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.
... We discuss how our findings correlate with recently published studies on survivin expression in gliomas. [27][28][29] ...
... This study demonstrates survivin protein expression and the presence of survivin mRNA in most gliomas and extends previous work by our group 33 and others. [27][28][29]34 We found that there was little variation in IHC staining intensity or percentage of positive tumor cells between our various GBM cases or between GBMs and lower-grade tumors. Reverse transcriptase-PCR allowed better quantitation and stratification of survivin levels in our gliomas than did IHC. ...
... Several recently published articles also have shown the presence of survivin in gliomas. [27][28][29] Our results are similar, but not identical, to these reports. ...
Context.—Survivin is a novel inhibitor of apoptosis that acts via a pathway independent of bcl-2. Little is known about its distribution in brain tumors or how it correlates with other biomarkers of malignancy, such as telomerase, an enzyme that plays a critical role in cellular immortalization and cancer biology.
Objectives.—To assess survivin protein expression in gliomas and to compare expression with that of telomerase.
Design.—Immunohistochemical staining for survivin protein expression was performed using an antibody developed in our laboratory. Quantitative survivin messenger RNA (mRNA) levels were assessed by reverse transcriptase-polymerase chain reaction. In selected cases, survivin results were compared with quantitative telomerase values analyzed by polymerase chain reaction–based telomerase repeat amplification protocol (TRAP) assay. Twenty-five tumor tissue samples from 16 cases of glioblastoma multiforme (GBM; including multiple tissue samples in 6 patients), 2 grade II gliomas, 4 grade III gliomas, and 3 control temporal lobectomy specimens were studied.
Results.—Nuclear immunoreactivity for survivin protein and survivin mRNA were detectable in most glioma samples, regardless of grade. Glioblastoma multiforme demonstrated moderate protein expression and survivin mRNA levels compared to epithelial malignancies previously tested in our laboratory. Although the association of survivin mRNA with the levels of telomerase within the GBM cases did not reach statistical significance, most GBMs also expressed survivin. The quantitative score for survivin mRNA was higher in GBMs than in grade II and III gliomas (P = .02), after accounting for multiple specimens per patient.
Conclusions.—Quantitative survivin mRNA analysis, but not immunohistochemistry, distinguished GBMs from lower grade gliomas. Mechanisms that promote both cell proliferation (telomerase expression) and cell survival (survivin expression) are often activated in GBMs.
... In this work, we explored whether GO would affect the immunogenicity of a known glioma peptide antigen (Ag). The peptide antigen is from the protein survivin, which is commonly expressed in human and murine malignant gliomas [20][21][22]. We found that a mixture of GO and Ag (GO-Ag) induced a more potent DC-mediated anti-glioma immune reaction in vitro. ...
... To induce a specific anti-glioma immune response, DCs must be exposed to glioma antigens. The antigen used in the study was a peptide (ELTLGEFLKL, termed Ag) from the protein survivin, which is widely expressed in malignant gliomas [20][21][22]. Survivin is a member of the inhibitor of apoptosis (IAP) protein family, which can regulate two important cellular processes: it inhibits apoptosis and promotes cell proliferation. Hence, survivin expression is generally associated with poor prognosis [30,31]. ...
... Hence, survivin expression is generally associated with poor prognosis [30,31]. The peptide ELTLGEFLKL can bind to HLA-A*0201, the most common human leukocyte antigen (HLA) serotype, and stimulate DCs to generate CD8 + immune responses against glioma cells [20][21][22]26]. Thus, the peptide was adopted in this study, mixed with GO at various feed ratios, and evaluated for its capability to induce a DC-mediated anti-glioma immune response. ...
Malignant glioma has extremely poor prognosis despite combination treatments with surgery, radiation, and chemotherapy. Dendritic cell (DC)-based immunotherapy may potentially serve as an adjuvant treatment of glioma, but its efficacy generally needs further improvement. Here we explored whether graphene oxide (GO) nanosheets could modulate the DC-mediated anti-glioma immune response in vitro, using the T98G human glioma cell line as the study model. Pulsing DCs with a glioma peptide antigen (Ag) generated a limited anti-glioma response compared to un-pulsed DCs. Pulsing DCs with GO alone failed to produce obvious immune modulation effects. However, stimulating DCs with a mixture of GO and Ag (GO-Ag) significantly enhanced the anti-glioma immune reaction (p < 0.05). The secretion of interferon gamma (IFN-γ) by the lymphocytes was also markedly boosted by GO-Ag. Additionally, the anti-glioma immune response induced by GO-Ag appeared to be target-specific. Furthermore, at the concentration used in this study, GO exhibited a negligible effect on the viability of the DCs. These results suggested that GO might have potential utility for boosting a DC-mediated anti-glioma immune response.
... The search strategy in Embase is as follows: ("glioblastoma"/exp OR "glioma"/exp OR glioblastoma OR glioma) AND ("immunotherapy"/exp OR immunotherapy OR (("therapy"/exp OR therapy) AND ("vaccine"/exp OR "vaccine" OR "immune checkpoint"/exp OR "immune checkpoint" OR "chimeric antigen receptor"/exp OR "chimeric antigen receptor" OR "microglia"/exp OR "microglia" OR "myeloid"))) AND The search strategy in Web of Science is as follows: (ALL = (glioblastoma OR glioma)) AND ((ALL = (immunotherapy)) OR ((ALL = (therapy)) AND ((ALL = (vaccine)) OR (ALL = (immune checkpoint)) OR (ALL = (chimeric antigen receptor)) OR (ALL = (microglia)) OR (ALL = (myeloid))))) AND 2002OR 2003OR 2004OR 2008OR 2009OR 2010OR 2011OR 2015 and English (Languages). ...
... Another frequently tumor-specific antigen is survivin (BIRC5). Although expressed during fetal development, survivin is rarely found in the healthy tissues of adult organisms [68]. SurVaxM is a long peptide mimic that spans amino acids 53 through 67 of the human survivin protein sequence, evaluated in the setting of phase II trial on newly diagnosed GBM (ndGBM) [16]. ...
... Survivin expression is increased in MDS [33] and MPN [64] relative to healthy controls and again peaks before malignant transformation but is not associated with survival [33]. Conversely, survivin expression is associated with faster progression of low-grade glioma to GBM [113] and diminished survival [20]. ...
... In spite of this difference, survivin offers a potential therapeutic target in both AML and GBM, as its expression in normal tissue is rare outside of fetal development [20]. Oligonucleotide inhibition of survivin has been shown by several authors to reduce cell viability and promote apoptosis in some AML cell lines [18,42,47] and, in combination with temozolomide, in GBM cell lines [23]. ...
Acute myeloid leukemia (AML) and glioblastoma (GB) are two malignancies associated with high incidence of treatment refractoriness and generally, uniformly poor survival outcomes. While the former is a hematologic (i.e. a “liquid”) malignancy and the latter a solid tumor, the two diseases share both clinical and biochemical characteristics. Both diseases exist predominantly in primary (de novo) forms, with only a small subset of each progressing from precursor disease states like the myelodysplastic syndromes or diffuse glioma. More importantly, the primary and secondary forms of each disease are characterized by common sets of mutations and gene expression abnormalities. The primary versions of AML and GB are characterized by aberrant RAS pathway, matrix metalloproteinase 9, and Bcl-2 expression, and their secondary counterparts share abnormalities in TP53, isocitrate dehydrogenase, ATRX, inhibitor of apoptosis proteins, and survivin that both influence the course of the diseases themselves and their progression from precursor disease. An understanding of these shared features is important, as it can be used to guide both the research about and treatment of each.
... Currently, the survivin protein has gained much attention for its role in tumor formation and cancer growth. Furthermore, survivin has attracted such great attention due to its high expression in a majority of cancers while being nearly undetectable in normal adult tissues [26,27]. Being a prominent member of the inhibitors of apoptosis (IAP) family, survivin plays an active role in allowing tumorous cells to avoid apoptosis while simultaneously increasing their rate of proliferation [28]. ...
... Many studies have focused on survivin due to its role and abilities, leading it to become a protein of high interest in the studies of cancer treatment as well as cancer immunization [29]. It has been reported that survivin expression has a clear prognostic value in human gliomas and that patients with survivin expression have significantly (p < 0.0001) shorter survival rates [27]. This review will predominately investigate the use of nanoparticle technology in treating cancers of the brain and for bypassing the BBB. ...
Brain cancer is a highly lethal disease, especially devastating toward both the elderly and children. This cancer has no therapeutics available to combat it, predominately due to the blood-brain barrier (BBB) preventing treatments from maintaining therapeutic levels within the brain. Recently, nanoparticle technology has entered the forefront of cancer therapy due to its ability to deliver therapeutic effects while potentially passing physiological barriers. Key nanoparticles for brain cancer treatment include glutathione targeted PEGylated liposomes, gold nanoparticles, superparamagnetic iron oxide nanoparticles and nanoparticle-albumin bound drugs, with these being discussed throughout this review. Recently, the survivin protein has gained attention as it is over-expressed in a majority of tumors. This review will briefly discuss the properties of survivin, while focusing on how both nanoparticles and survivin-targeting treatments hold potential as brain cancer therapies. This review may provide useful insight into new brain cancer treatment options, particularly survivin inhibition and nanomedicine.
... The expression of survivin has been significantly correlated with the malignant grade of astrocytic tumors and a worse clinical outcome (16). Furthermore, patients who had been diagnosed with glioblastomas, with over-expression of survivin, have been observed to have a significantly shorter survival time (17). Therefore, it is suggested that survivin may be involved in tumorigenesis and malignant progression of glial cell tumors (18). ...
... Various previous studies have shown a correlation between survivin expression and radiation resistance in certain human cancers. Chakravarti et al (17) reported that survivin improves double-strand DNA break repair in tumor cells, inhibiting radiation-induced cell death. Sasaki et al (15) demonstrated that suppression of survivin by small interfering ribonucleic acid improved the radiosensitivity of tumor cells. ...
Apoptosis, whose mechanism remains unclear, is regulated by multiple factors. B-cell lymphoma 2 (Bcl-2) is a well-known anti-apoptotic mediator. Survivin is also a recently recognized novel family inhibitor of apoptosis protein, which inhibits apoptosis via a pathway distinct from Bcl-2 family members. Survivin and Bcl-2 are expressed in various types of human cancer. In the present study, survivin and Bcl-2 expression were characterized in glial cell tumors, and the correlation with pathological malignancy and anti-apoptotic properties were investigated. Fifty-eight patients who had undergone surgical resection for glial cell tumors were evaluated. The pathological types of glial cell tumors were categorized according to the World Health Organization classification. Survivin and Bcl-2 expression levels were investigated by western blot analysis, and apoptosis was detected by DNA fragmentation analysis. The anti-apoptotic rate of glial cell tumors was calculated in tumor samples according to the expression of survivin and Bcl-2 or co-expression. Survivin was characterized in 60.3%, and Bcl-2 was expressed in 43.1% of glioma samples. Co-expression of survivin and Bcl-2 was observed in 25.9% of the tumor specimens. Survivin expression in astrocytic tumors was identified to be significantly associated with the pathological grade (P<0.05); however, Bcl-2 was not (P>0.05). Anti-apoptotic rate of glial cell tumors were detected in 91.4, 92.0 and 100% of patients exhibiting survivin, Bcl-2 or co-expression, respectively. However, the difference in anti-apoptotic frequency between the three groups was not identified to be statistically significant (P>0.05). The present study suggests that survivin expression is correlated with pathological grades of gliomas. In addition, the expression of survivin or Bcl-2 exerts potent anti-apoptotic properties in gliomas. Thus, survivin or Bcl-2 may serve as potential targets for inducing the apoptosis of gliomas.
... Therefore, their expression in cancers is usually correlated with poor prognosis. 7,8 IAPs are characterized by the presence of one to three Baculovirus IAP Repeat (BIR) domains, necessary for protein-protein interactions like caspase inhibitory interactions, typically arranged in the protein's amino terminus. cIAP1, cIAP2, XIAP and ML-IAP also contain a carboxylterminus RING (Really Interesting New Gene) domain, working as an E3-ubiquitin ligase. ...
... 10 For patients with GBM, the prognostic value of cIAP1, cIAP2, XIAP and ML-IAP expression remains to be determined. 7,11 Therefore, in this study we investigated ex vivo the correlation between their expression levels and patient's survival in two retrospective cohorts in order to highlight the most interesting druggable targets. In order to assess the effect of SMAC mimetic on GBM tumors, we then tested in vitro the effect of GDC-0152 on four GBM cell lines. ...
Glioblastomas (GBMs) are the most aggressive primary brain tumors in adult and remain a therapeutic challenge. Targeting key apoptosis regulators with the ultimate aim to restore apoptosis in tumor cells could be an interesting therapeutic strategy. The inhibitors of apoptosis proteins (IAPs) are regulators of cell death and represent attractive targets, especially because they can be antagonized by SMAC mimetics. In this study, we first investigated the expression of cIAP1, cIAP2, XIAP and ML-IAP in human GBM samples and in four different cell lines. We showed that all GBM samples and GBM cell lines expressed all these IAPs, although the expression of each IAP varied from one case to another. We then showed that high level of ML-IAP predicted worse progression-free survival and overall survival in both univariate and multivariate analyses in two independent cohorts of 58 and 43 primary human GBMs. We then used GDC-0152, a SMAC mimetic that antagonizes these IAPs and confirmed that GDC-0152 treatment in vitro decreased IAPs in all the cell lines studied. It affected cell line viability and triggered apoptosis, although the effect was higher in U87MG and GL261 than in GBM6 and GBM9 cell lines. In vivo, GDC-0152 effect on U87MG orthotopic xenografts was dose dependent; it postponed tumor formation and slowed down tumor growth, significantly improving survival of GBM-bearing mice. This study revealed for the first time that ML-IAP protein expression correlates with GBM patient survival and that its antagonist GDC-0152 improves outcome in xenografted mouse.
... Survivin is an inhibitor of the apoptotic protein (IAP) that plays an important role in apoptosis inhibition and promotion of the G 2 /M phase cell cycle transition that is essential for cancer progression (6,35,49,59). Survivin is highly expressed in GMB cells in comparison with normal brain cells and it is related to GBM patients with poor prognosis and apoptosis resistance (8,19). Inhibition of survivin activity is considered as a crucial strategy in GBM treatment (21). ...
... Inhibition of mTOR may trigger autophagy process in apoptosis-resistant cancer cells through blockage of the Akt/mTOR pathway (4). Akt is similar to survivin in being highly expressed in GBM cells, and in its correlation with invasive activities of GBM (8). Several studies have discussed the possibility that interactions between autophagy and simultaneous inhibition of Akt and survivin can enhance GBM autophagic cell death (1). ...
Chemotherapy efficacy is limited by intrinsic and acquired resistance in glioblastoma (GBM); hence, novel tactics are crucial. Survivin has been demonstrated as a key resistant factor in GBM because of its function in inhibiting apoptosis, regulating autophagy, and in promoting G₂/M cell cycle transition. Parthenolide has been reported to be an effective antitumor agent in a variety of tumor cells and decreases survivin level in leukemia cells. But the effect of parthenolide on survivin and the cell death process in GBM is still unknown. The aim of this study was to examine whether parthenolide had the potential to inhibit cell proliferation in the GBM cell line U373. The parthenolide-induced effects in relation to survivin suppression and cell death were further investigated. Our results showed that parthenolide substantially inhibited cell viability with IC₅₀ values of approximate 16 μM. Treatment with parthenolide at the dose of 16 μM led to considerable downregulation of survivin, G₂/M cell cycle arrest and Chk2 upregulation in cells. Parthenolide induced apoptosis in only a few cells and a slight increase in activated caspases 3 levels. By contrast, parthenolide induced a significant increase of intracellular autophagosomes and the expression of autophagy related proteins, including ULK1 and LC3 I/LC3 II, in the treated cells. These results suggested that parthenolide induced survivin inhibition, G₂/M cell cycle arrest, and triggered cell death through autophagic cell death in the GBM cell line.
... The expression of survivin (BIRC5), a member of the inhibitor of apoptosis gene family, has been shown to correlate negatively with survival in patients with high-grade gliomas. 7,8 Survivin transcript variants can be pro-or anti-apoptotic and are differentially expressed in various cancers. In addition to having unique biological activities, survivin splice variants appear to correlate with different clinical outcomes in other types of cancer. ...
... In GBM, increased protein levels of survivin from human tumor samples have been shown to correlate negatively with overall survival. 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. ...
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.
... Higher survivin expression in patients with glioma has been associated with resistance to temozolomide therapy and worse overall survival. 31,32 Furthermore, syndecan-1 expression in human glioma is associated with advanced tumour progression and poor prognosis. 33 The decrease in survivin and syndecan-1 expression following NSC-CRAd-S-pk7 therapy might be due to the replication of CRAd-S-pk7 in the glioma cells, which leads to programmed cell death. ...
Background
Malignant glioma is the most common and lethal primary brain tumour, with dismal survival rates and no effective treatment. We examined the safety and activity of NSC-CRAd-S-pk7, an engineered oncolytic adenovirus delivered by neural stem cells (NSCs), in patients with newly diagnosed high-grade glioma.
Methods
This was a first-in-human, open-label, phase 1, dose-escalation trial done to determine the maximal tolerated dose of NSC-CRAd-S-pk7, following a 3 + 3 design. Patients with newly diagnosed, histologically confirmed, high-grade gliomas (WHO grade III or IV) were recruited. After neurosurgical resection, NSC-CRAd-S-pk7 was injected into the walls of the resection cavity. The first patient cohort received a dose starting at 6·25 × 1010 viral particles administered by 5·00 × 107 NSCs, the second cohort a dose of 1·25 × 1011 viral particles administered by 1·00 × 108 NSCs, and the third cohort a dose of 1·875 × 1011 viral particles administered by 1·50 × 108 NSCs. No further dose escalation was planned. Within 10–14 days, treatment with temozolomide and radiotherapy was initiated. Primary endpoints were safety and toxicity profile and the maximum tolerated dose for a future phase 2 trial. All analyses were done in all patients who were included in the trial and received the study treatment and were not excluded from the study. Recruitment is complete and the trial is finished. The trial is registered with ClinicalTrials.gov, NCT03072134.
Findings
Between April 24, 2017, and Nov 13, 2019, 12 patients with newly diagnosed, malignant gliomas were recruited and included in the safety analysis. Histopathological evaluation identified 11 (92%) of 12 patients with glioblastoma and one (8%) of 12 patients with anaplastic astrocytoma. The median follow-up was 18 months (IQR 14–22). One patient receiving 1·50 × 108 NSCs loading 1·875 × 1011 viral particles developed viral meningitis (grade 3) due to the inadvertent injection of NSC-CRAd-S-pk7 into the lateral ventricle. Otherwise, treatment was safe as no formal dose-limiting toxicity was reached, so 1·50 × 108 NSCs loading 1·875 × 1011 viral particles was recommended as a phase 2 trial dose. There were no treatment-related deaths. The median progression-free survival was 9·1 months (95% CI 8·5–not reached) and median overall survival was 18·4 months (15·7–not reached).
Interpretation
NSC-CRAd-S-pk7 treatment was feasible and safe. Our immunological and histopathological findings support continued investigation of NSC-CRAd-S-pk7 in a phase 2/3 clinical trial.
Funding
US National Institutes of Health.
... It has been reported that survivin expression may provide prognostic information in various types of cancers, such as esophageal cancer (Kato et al., 2001;Pennati et al., 2007), breast cancer (Altieri, 2003), lung cancer (Velculescu et al., 1999;Xu et al., 2012), gastric cancer (Yamamoto and Tanigawa, 2001), colorectal cancer (Waligorska-Stachura et al., 2012), and glioma (Liang et al., 2017), and patients with high survivin expression had significantly shorter survival rates. Therefore, survivin has been intensely pursued as a target for cancer therapy alone or in combination with other therapeutic strategies (Kato et al., 2001;Chakravarti et al., 2002;Wang et al., 2007). ...
Efforts have been made to find effective medical drugs for cervical cancer treatment. The incidence of cervical cancer ranks second among women, and is a serious threat to women's health. Aberrant activation of the nonreceptor protein tyrosine kinases such as Src is commonly observed in progressive stages of human tumors. Thus, targeting Src kinase could be a promising strategy for cervical cancer therapy. In this study, we explored the potential utility of bosutinib in the treatment of cervical cancer. We found that bosutinib, as a potent dual Src/Abl inhibitor, could exert anti‐tumor effects on cervical cancer. Bosutinib inhibited cervical cancer cells proliferation and colony formation ability in a dose‐dependent manner, and also induced apoptosis. Mechanistically, bosutinib effectively decreased the activity of Src/NF‐κB/survivin signaling pathway. Our study provided a biological rationale to test bosutinib as a valuable therapeutic option for cervical cancer patients. Anat Rec, 302:2193–2200, 2019. © 2019 American Association for Anatomy
... Survivin immunoreactivity was upregulated significantly in high-grade glioma, but it was not correlated with gender, age, tumor size or location. In the follow-up data, patients with survivin-positive tumors experienced a significantly shorter survival time compared with those who were negative for survivin (16 vs. 24 months), and this was in agreement with previous studies (31,32). Furthermore, survivin may be used to predict prognosis for low-grade and high-grade gliomas. ...
The present study aimed to determine cyclooxygenase 2 (COX-2) and survivin expression levels in glioma tissues, and to investigate their association with clinicopathological factors and patient survival. Immunohistochemistry was performed to evaluate COX-2 and survivin expression levels in paraffin-embedded surgically resected tissues from 70 patients with glioma and 7 individuals with normal brain tissues. The association between COX-2 and survivin expression levels and clinicopathological features was investigated using the χ²test, and the survival time was analyzed using the Kaplan Meier method with log-rank test. COX-2 and survivin were overexpressed in glioma tissues, and higher expression levels were observed in glioma tissues of histological grades III-IV compared with those in grade I-II tumor tissues (P0.05). There was a significant positive association between the expression levels of COX-2 and survivin in the glioma tissues. Additionally, COX-2 and survivin expression levels were significantly negatively correlated with the rate of survival. In conclusion, COX-2 and survivin expression is positively associated with the pathological grade of a glioma and may contribute to glioma tumorigenesis. Therefore, COX-2 and survivin may be sensitive predictors of a negative clinical prognosis for patients with glioma.
... Increasing efforts are made to improve HGG treatment. Recently, several studies have focused on using gene expression profiles to identify potential new biomarkers for diagnosis, prognosis, staging and therapy development [7,8]. Multiple studies suggest that gene expression-based classification of malignant gliomas may correlate better with survival than histological classification [9], and provide a useful method to identify previously unrecognized but clinically relevant prognostic indicators [10,11]. ...
Current treatment methods for patients diagnosed with gliomas have shown limited success. This is partly due to the lack of prognostic genes available to accurately predict disease outcomes. The aim of this study was to investigate novel prognostic genes based on the molecular profile of tumor samples and their correlation with clinical parameters. In the current study, microarray data (GSE4412 and GSE7696) downloaded from Gene Expression Omnibus were used to identify differentially expressed prognostic genes (DEPGs) by significant analysis of microarray (SAM) between long-term survivors (>2 years) and short-term survivors (≤2 years). DEPGs generated from these two datasets were intersected to obtain a list of common DEPGs. The expression of a subset of common DEPGs was then independently validated by real-time reverse transcription quantitative PCR (qPCR). Survival value of the common DEPGs was validated using known survival data from the GSE4412 and TCGA dataset. After intersecting DEPGs generated from the above two datasets, three genes were identified which may potentially be used to determine glioma patient prognosis. Independent validation with glioma patients tissue (n = 70) and normal brain tissue (n = 19) found PPIC, EMP3 and CHI3L1 were up-regulated in glioma tissue. Survival value validation showed that the three genes correlated with patient survival by Kaplan-Meir analysis, including grades, age and therapy.
... Finally, these findings were associated with the decreased expression of caspase-9 inhibitors Xiap and survivin (26). Survivin down-regulation should have therapeutic importance, since its expression is found to be associated with the drug-resistant melanoma phenotype (27) and it is inversely correlated with the survival rate of glioma patients (28). ...
Marrubium vulgare is a European medicinal plant with numerous beneficial effects on human health. The aim of the study was to isolate the plant ethanolic extract (MVE) and to investigate its anti-melanoma and anti-glioma effects. MVE was prepared by the modified pharmacopoeial percolation method and characterized by UHPLC-LTQ OrbiTrap MS. MVE dose-dependently reduced viability of melanoma (B16) and glioma (U251) cells, but not peripheral blood mononuclear cells. It arrested cell cycle in S+G2/M phase, which was associated with the activation of MAP kinase p38 and up-regulation of antiproliferative genes p53, p21 and p27. MVE induced oxidative stress, while antioxidants abrogated its antitumor effect. Furthermore, MVE induced mitochondrial depolarization, activation of caspase-9 and -3, Parp cleavage, phosphatidylserine exposure and DNA fragmentation. The mitochondrial apoptotic pathway was associated with the up-regulation of proapoptotic genes Pten, Bak1, Apaf1, and Puma and down-regulation of antiapoptotic genes survivin and Xiap. MVE also stimulated the expression of autophagy-related genes Atg5, Atg7, Atg12, Beclin-1, Gabarab and Sqstm1, as well as LC3-I conversion to the autophagosome associated LC3-II, while autophagy inhibitors exacerbated its cytotoxicity. Finally, the most abundant phenolic components of MVE, ferulic, p-hydroxybenzoic, caffeic and chlorogenic acids, did not exert a profound effect on viability of tumor cells, suggesting that other components individually or in concert are the mediators of the extracts' cytotoxicity. By demonstrating the ability of MVE to inhibit proliferation, induce apoptosis and cytoprotective autophagy, our results suggest that MVE, alone or combined with autophagy inhibitors, could be a good candidate for anti-melanoma and anti-glioma therapy.
... However, patient sample 2 (PS2) showed only a marginal increase in the Survivin expression. The increased expression of Survivin in the recurrent cells formed upon lethal exposure of radiation is noteworthy since Survivin has been shown to be overexpressed in Glioblastoma tumours associated with high anti-apoptotic activity 36,37 . The expression of other pro-survival gene Bcl-xL in the recurrent cells remain unaltered except for SF268 cells (Fig. 2b), may be due to the dependency of GBM recurrent cells on Survivin for its anti-apoptotic activity. ...
An inability to discern resistant cells from bulk tumour cell population contributes to poor prognosis in Glioblastoma. Here, we compared parent and recurrent cells generated from patient derived primary cultures and cell lines to identify their unique molecular hallmarks. Although morphologically similar, parent and recurrent cells from different samples showed variable biological properties like proliferation and radiation resistance. However, total RNA-sequencing revealed transcriptional landscape unique to parent and recurrent populations. These data suggest that global molecular differences but not individual biological phenotype could differentiate parent and recurrent cells. We demonstrate that Raman Spectroscopy a label-free, non-invasive technique, yields global information about biochemical milieu of recurrent and parent cells thus, classifying them into distinct clusters based on Principal-Component-Analysis and Principal-Component-Linear-Discriminant-Analysis. Additionally, higher lipid related spectral peaks were observed in recurrent population. Importantly, Raman spectroscopic analysis could further classify an independent set of naïve primary glioblastoma tumour tissues into non-responder and responder groups. Interestingly, spectral features from the non-responder patient samples show a considerable overlap with the in-vitro generated recurrent cells suggesting their similar biological behaviour. This feasibility study necessitates analysis of a larger cohort of naïve primary glioblastoma samples to fully envisage clinical utility of Raman spectroscopy in predicting therapeutic response.
... Survivin has been reported to be abundantly overexpressed in malignant gliomas [29]. Our study showed that survivin immune-expression was different significantly among various grades of astrocytoma (P < 0.001). ...
... Survivin is well known for its anti apoptotic function and has been reported to be upregulated in many cancers, including gliomas [14][15][16][17][18][19][20]. A survivin luciferase reporter assay in rat neurons and various glioma cells confirmed overexpression of the gene in glioma cells (Figure 2A). ...
Oncolytic herpes simplex virus type 1 (oHSV-1) therapy is an emerging treatment modality that selectively destroys cancer. Here we report use of a glioma specific HSV-1 amplicon virus (SU4-124 HSV-1) to selectively target tumour cells. To achieve transcriptional regulation of the SU4-124 HSV-1 virus, the promoter for the essential HSV-1 gene ICP4 was replaced with a tumour specific survivin promoter. Translational regulation was achieved by incorporating 5 copies of microRNA 124 target sequences into the 3'UTR of the ICP4 gene. Additionally, a 5'UTR of rat fibroblast growth factor -2 was added in front of the viral ICP4 gene open reading frame. Our results confirmed enhanced expression of survivin and eIF4E in different glioma cells and increased micro-RNA124 expression in normal human and mouse brain tissue. SU4-124 HSV-1 had an increased ICP4 expression and virus replication in different glioma cells compared to normal neuronal cells. SU4-124 HSV-1 exerted a strong antitumour effect against a panel of glioma cell lines. Intracranial injection of SU4-124 HSV-1 did not reveal any sign of toxicity on day 15 after the injection. Moreover, a significantly enhanced antitumour effect with the intratumourally injected SU4-124 HSV-1 virus was demonstrated in mice bearing human glioma U87 tumours, whereas viral DNA was almost undetectable in normal organs. Our study indicates that incorporation of multiple cancer-specific regulators in an HSV-1 system significantly enhances both cancer specificity and oncolytic activity.
... We have shown previously that suppression of apoptosis can promote malignant progression in a murine model of glioma. Increased expression of Survivin (BIRC5), a member of the Inhibitors of Apoptosis (IAP) family, has been shown to be prognostic of shorter survival in patients with HGG [3,4]. We showed that the overexpression of anti-apoptotic Survivin transcript variant 2 is associated with a poor prognosis as well as increased angiogenesis in a murine model [5]. ...
Gliomas, the most common primary brain tumor in humans, include a spectrum of disease. High-grade gliomas (HGG), such as glioblastoma, may arise from low-grade gliomas (LGG) that have a more indolent course. The process of malignant transformation (MT) of LGG to HGG is poorly understood but likely involves the activation of signaling programs that suppress apoptosis. We previously showed that Survivin (BIRC5) plays a role in malignant progression of glioma. Here, we investigated the role of the remaining members of the Inhibitors of Apoptosis (IAP) family on promoting MT in glioma. Utilizing expression data from the cancer genome atlas (TCGA), we identified BIRC3 as a key facilitator of MT from LGG to HGG. TCGA HGGs with high expression of BIRC 3 demonstrated a survival disadvantage and expression levels of BIRC3 were also significantly higher in TCGA HGG compared to TCGA LGG cases. We validated our findings from TCGA by using matched human specimens to show that BIRC expression is increased in HGG compared to their precursor LGG lesions. Using a unique murine model of glioma, we show that overexpression of BIRC3 promotes higher grade glioma and significantly reduces tumor-free survival in mice.
... [22][23] . 一 些研究表明, s u r v i v i n的高表达与肿瘤的预后 差和耐药性有关 [24][25][26] . survivin的过表达与胰腺 癌的发生、发展密切相关 [27] . ...
AIM: To investigate the therapeutic effects of survivin antisense oligonucleotide (ASODN) on nude mice bearing human pancreatic carcinoma xenograft. METHODS: The nude mouse model of pancreatic cancer was established using human pancreatic cancer cell line BxPC-3. The mice bearing tumor were intratumorally injected with survivin ASODN (40 g/200 L per mouse). The tumor size and volume were measured before and after injection. The pathological changes of tumor tissues were observed. The expression level of survivin mRNA was measured by reverse transcription-polymerase chain reaction (RT-PCR), and the activity of caspase-3 was evaluated using a caspase-3 assay kit. The expression of proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) were detected by immunohistochemistry. RESULTS: Twenty days after injection, the tumor volume and weight were significantly decreased in survivin ASODN group as compared with those in the control and SODN group (427.34 ± 12.44 mm3 vs 703.56 ± 12.51, 687.59 ± 12.44 mm3, P < 0.01; 0.57 ± 0.06 g vs 1.16 ± 0.12, 1.07 ± 0.10 g, P < 0.01), and the tumor inhibition rate was 50.86%. The expression of survivin mRNA was decreased by 50%, while the caspase-3 activity was significantly higher in survivin ASODN group than that in the control and SODN group (0.040 ± 0.018 vs 0.006 ± 0.001, 0.007 ± 0.002, P < 0.01). The expression of PCNA and MVD were significantly lower in survivin ASODN group than those in the control and SODN group (28.33 ± 2.16 vs 35.17 ± 3.71, 34.33 ± 3.27, P < 0.01; 15.50 ± 3.08 vs 21.33 ± 2.94, 20,67 ± 2.16, P < 0.01). However, both expression of PCNA and MVD had no significant difference between the control and SODN group (P > 0.05). CONCLUSION: Survivin ASODN can inhibit the growth of human pancreatic carcinoma xenograft in nude mouse through activating caspase-3 to induce apoptosis, and suppressing the proliferation of pancreatic cancer cells as well as the angiogenesis of carcinoma.
... Apoptosis, like cellular proliferation, is under very tight genetic controls. The loss of normal regulation of apoptosis has been implicated in the development and progression of gliomas in a number of studies (Alderson, 1995;Sano et al., 1999;Rodriguez-Pereira, 2001;Chakravarti, 2002). We shall briefly discuss some of genes found to be implicated in the control of cellular proliferation and apoptosis in human gliomas. ...
Gliomas are the most common intracranial malignant tumors in humans, and high-grade gliomas in particular pose a unique challenge due to their propensity for proliferation and tissue invasion. Our understanding of glioma oncogenesis, proliferation, and invasion has been greatly advanced in the past 10 years as researchers have gained a better understanding of the molecular biology of these tumors. This article highlights glioma histopathology, as well as cytogenetic and molecular alterations associated with the pathogenesis of human gliomas. It is hoped that better understanding of the molecular pathogenesis of gliomas will improve tumor classification as well as lead to novel targets for therapy and prognostic markers.
... Inhibitor of apoptosis family proteins (IAPs), which can bind to and inhibit numerous caspases, have also been observed to be deregulated through elevated expression in GBM [16]. One the most well characterized IAPs in GBM, survivin, is inversely correlated with overall survival and is associated with reduced apoptotic capacity [17]. Inhibition of IAP proteins has been an effective strategy to overcome apoptotic resistance in response to a variety of therapeutic strategies including growth factor inhibitors [18], radiotherapy [19], Apo2L/TRAIL [20], and chemotherapy with Temozolomide (TMZ) [21]. ...
The five-year survival rate for patients with malignant glioma is less than 10 %. Despite aggressive chemo/radiotherapy these tumors have remained resistant to almost every interventional strategy evaluated in patients. Resistance to these agents is attributed to extrinsic mechanisms such as the tumor microenvironment, poor drug penetration, and tumoral heterogeneity. In addition, genetic and molecular examination of these tumors has revealed defective apoptotic regulation, enhanced pro-survival autophagy signaling, and a propensity for necrosis that aids in the adaptation to environmental stress and resistance to treatment. The combination of extrinsic and intrinsic hallmarks in glioma contributes to the multifaceted resistance to traditional anti-tumor agents. Here we describe the biology of the disease relevant to therapeutic resistance, with a specific focus on molecular deregulation of cell death pathways. Emerging studies investigating the targeting of these pathways including BH3 mimetics and autophagy inhibitors that are being evaluated in both the preclinical and clinical settings are discussed. This review highlights the pathways exploited by glioblastoma cells that drive their hallmark pro-survival predisposition and makes therapy development such a challenge.
... Specifically, its anti-apoptotic function seems to be related to the ability to directly or indirectly inhibit caspases. Survivin is expressed in embryonic tissues as well as in the majority of human cancers, but is not expressed in most normal adult tissues and is associated with chemotherapy resistance (34), and patients with survivin-expressing Table V tumours have poorer survival (35). Studies have found that patients with tumours that do not express survivin or only express it in low levels have more favourable outcomes. ...
We studied the effect of resveratrol treatment on multidrug-resistant human non-small cell lung cancer cells. Human multidrug-resistant SPC-A-1/CDDP cells were treated with resveratrol at a concentration of 25, 50, or 100 μM in in vitro studies and nude mice were implanted with multidrug-resistant SPC-A-1/and fed a special diet that included resveratrol at a dose of either 1 g/kg/day or 3 g/kg/ day in in vivo studies. No adverse toxicological effects of resveratrol treatment were observed. The rate of cell proliferation, apoptosis ratio, cell cycle phase distribution, IC 50 values of cisplatin, gefitinib, and paclitaxel, implanted tumour volume, and expression of survivin in resveratrol-treated and control mice were then determined. Resveratrol significantly inhibited the proliferation of SPC-A-1/CDDP cells, induced apoptosis, arrested the cell cycle phase between G 0 -G 1 and S phase or at the G 2 /M phase, decreased the IC 50 values of multiple chemotherapeutic drugs, and showed anti-tumour effects in nude mice that had been implanted with SPC-A-1/ CDDP cells. In additional, resveratrol affected the proliferation of SPC-A-1/CDDP cells in a dose- and time-dependent manner. Expression of survivin in SPC-A-1/CDDP cells decreased after they were treated with all concentrations of resveratrol and resveratrol was also found to have a dose-dependent effect on survivin expression. Resveratrol can induce apoptosis in multidrug-resistant human NSCLC SPC-A-1/ CDDP cells by down-regulating the expression of survivin.
... However in a recent phase III clinical trial, it was reassessed the effect in GBM from patients treated with TTFields against the outcomes from patients treated with standard chemotherapy [8]. In this study, no benefit was observed with TTFields in comparison to chemotherapy, despite the fact that that chemotherapy has a very low effectiveness in the treatment of GBM [9,10]. Nevertheless, researchers that promote TTFields point to the fact that patients get a better quality of life for the same treatment outcome. ...
Delivery of the so-called Tumor Treatment Fields (TTFields) has been proposed as a cancer therapy. These are low magnitude alternating electric fields at frequencies from 100 to 300kHz which are applied continuously in a non-invasive manner. Electric field delivery may produce an increase in temperature which cannot be neglected. We hypothesized that the reported results obtained by applying TTFields in vivo could be due to heat rather than to electrical forces as previously suggested. Here, an in vivo study is presented in which pancreatic tumors subcutaneously implanted in nude mice were treated for a week either with mild hyperthermia (41°C) or with TTFields (6V/cm, 150kHz) and tumor growth was assessed. Although the TTFields applied singly did not produce any significant effect, the combination with chemotherapy did show a delay in tumor growth in comparison to animals treated only with chemotherapy (median relative reduction=47%). We conclude that concomitant chemotherapy and TTFields delivery show a beneficial impact on pancreatic tumor growth. Contrary to our hypothesis, this impact is non-related with the induced temperature increase.
Copyright © 2015 Elsevier B.V. All rights reserved.
... In addition, survivin also plays an important role in the angiogenesis process. Overexpression of survivin in gliomas is significantly associated with tumor genesis and progression, as well as with poor prognosis of patients (43,44). Recently, Virrey et al (45) demonstrated that overexpression of survivin in endothelial cells serves as a protective mechanism that defends the vasculature from drug cytotoxicity. ...
According to World Health Organization criteria, diffuse gliomas are divided into several histological subtypes, including astrocytomas, oligodendrogliomas, and oligoastrocytomas, and 4 malignancy grades (I-IV). Molecular alterations, such as the isocitrate dehydrogenase gene (IDH) mutation or 1p/19q loss, are found in these tumors but are not included in the current classification system. Recently, mutation of α thalassemia/mental retardation syndrome X-linked (ATRX) gene and its loss of expression have been reported in infiltrating gliomas. We evaluated ATRX protein expression in 272 gliomas and its association with molecular and clinical features. Loss of ATRX expression was more common in tumors with an astrocytic component (astrocytomas II/III, 46.4%; oligoastrocytomas, 47.5%) but was uncommon in oligodendrogliomas (7.3%) and glioblastomas (0.9%). In astrocytic tumors, loss of ATRX expression was significantly associated with longer overall survival. Remarkably, on the basis of IDH mutation, 1p/19q codeletion, and ATRX expression, our study defined 4 molecularly and prognostically different groups of gliomas, showing the relevance of ATRX expression as a new marker for refining the molecular classification of gliomas and for distinguishing clinically distinct prognostic subgroups of patients.
... CTGF was proved to be a prognostic for tumor progression and survival of gliomas [24], and targeting CTGF was correlated with improved survival of glioblastoma [25]. Survivin expression levels also are of prognostic values in human gliomas [26], and high expression of survivin enhances radiation resistance in glioblastoma [27]. Moreover, both CTGF and survivin are involved in the regulation of cancer stem cell self-renewal [11,27]. ...
... CRAd-S-pk7 is a replication-competent adenoviral vector that contains the wild-type adenovirus replication protein, E1A, under the control of the human survivin promoter, a gene that is known to be overexpressed in malignant glioma. 24,25 This viral vector was created via homologous recombination using a shuttle plasmid containing the survivin promoter upstream of the E1A gene and an adenoviral vector backbone modified to contain a poly-lysine (pk7) addition onto the C terminus of the wild-type fiber protein. 18 To load HB1.F3.CD cells, cell suspensions were incubated with DMEM containing 10% fetal bovine serum and 50 IU per cell of CRAd-Survivin-pk7 viruses. ...
In preclinical studies, neural stem cell (NSC)-based delivery of oncolytic virus has shown great promise in the treatment of malignant glioma. Ensuring the success of this therapy will require critical evaluation of the spatial distribution of virus after NSC transplantation. In this study, the patient-derived GBM43 human glioma line was established in the brain of athymic nude mice, followed by the administration of NSCs loaded with conditionally replicating oncolytic adenovirus (NSC-CRAd-S-pk7). We determined the tumor coverage potential of oncolytic adenovirus by examining NSC distribution using magnetic resonance (MR) imaging and by three-dimensional reconstruction from ex vivo tissue specimens. We demonstrate that unmodified NSCs and NSC-CRAd-S-pk7 exhibit a similar distribution pattern with most prominent localization occurring at the tumor margins. We were further able to visualize the accumulation of these cells at tumor sites via T2-weighted MR imaging as well as the spread of viral particles using immunofluorescence. Our analyses reveal that a single administration of oncolytic virus-loaded NSCs allows for up to 31% coverage of intracranial tumors. Such results provide valuable insights into the therapeutic potential of this novel viral delivery platform.Cancer Gene Therapy advance online publication, 19 December 2014; doi:10.1038/cgt.2014.72.
... We will address more differences between autologous and allogeneic sources of tumor antigens later (Table 1). An advantage of whole tumor cell lysates is the ability to utilize multiple tumor associated antigens (TAA) such as GnT-V, 22 survivin, 23 IL13Ra2, 24 tyrosinase-related protein 1 (Trp-1) and Trp-2, 25,26 Gage, 27 human melanoma-associated antigens p97/ GP100, 10 EphA2/Eck, 28 Aim-2, 29 Sart-1, 30 Her2/ nu 31 and Mage-1 31 to elicit an immune response. 32 Although vaccines consisting of whole cell tumor lysates have shown some signs of successful therapy, they failed to receive FDA approval in the United States. ...
Anti-tumor immunotherapy using tumor lysate-based vaccines has made great advances over recent decades. Cancer vaccines aim to elicit adaptive immune responses through various pathways by providing tumor and tumor-associated antigens with an immune stimulant or adjuvant. These anti-tumor vaccines are therefore developed as personalized treatments. Utilizing tumors as a source of vaccine antigens in immunotherapy has demonstrated promising results with minimal toxicity. However, to date, researchers have failed to overcome the overpowering immune suppressive effects within the tumor microenvironment. Immune suppression occurs naturally via multiple mechanisms. These mechanisms serve an important homeostatic role restoring a normal tissue microenvironment following an inflammatory response. Due to these suppressive mechanisms and the inherent heterogeneity of tumors, it is imperative to then elicit and maintain a specific tumoricidal response if vaccine therapy or some other combination of reagents is chosen. In this review, we focus on the historical use of tumors as a source of antigens to elicit a tumoricidal response and the limitations encountered that prevent greater success in immunotherapy. We describe the advantages and disadvantages of various vaccines and their ineffectiveness due to tumor-induced immune suppression.
... IAP family of proteins is known to act as caspase inhibitors, blocking caspase activation and further inhibiting apoptosis (9). Previous studies have shown that a number of different IAPs were highly expressed in malignant gliomas including XIAP (23), apollon (24), and survivin (25), with an adverse correlation with patient outcomes. Although less studies are carried out in brain tumors, recent studies reported that Livin and survivin are upregulated in glioma stem cells (GSC) derived from human glioblastoma and astrocytoma tissues or glioblastoma cell lines but are the most increased in just differentiated GSC (26,27). ...
Purpose:
Tumor hypoxia is one of the crucial microenvironments to promote therapy resistance (TR) in glioblastoma multiforme (GBM). Livin, a member of the family of inhibitor of apoptosis proteins, contributes antiapoptosis. However, the role of tumor hypoxia in Livin regulation and its impact on TR are unclear.
Experimental design:
Livin expression and apoptosis for tumor hypoxic cells derived from human glioblastoma xenografts or in vitro hypoxic stress-treated glioblastoma cells were determined by Western blotting, immunofluorescence imaging, and annexin V staining assay. The mechanism of hypoxia-induced Livin induction was investigated by chromatin immunoprecipitation assay and reporter assay. Genetic and pharmacologic manipulation of Livin was utilized to investigate the role of Livin on tumor hypoxia-induced TR in vitro or in vivo.
Results:
The upregulation of Livin expression and downregulation of caspase activity were observed under cycling and chronic hypoxia in glioblastoma cells and xenografts, concomitant with increased TR to ionizing radiation and temozolomide. However, knockdown of Livin inhibited these effects. Moreover, hypoxia activated Livin transcription through the binding of hypoxia-inducible factor-1α to the Livin promoter. The targeted inhibition of Livin by the cell-permeable peptide (TAT-Lp15) in intracerebral glioblastoma-bearing mice demonstrated a synergistic suppression of tumor growth and increased the survival rate in standard-of-care treatment with radiation plus temozolomide.
Conclusions:
These findings indicate a novel pathway that links upregulation of Livin to tumor hypoxia-induced TR in GBM and suggest that targeting Livin using cell-permeable peptide may be an effective therapeutic strategy for tumor microenvironment-induced TR.
... TRP-2-specific CTL responses have been detected in GBM patients vaccinated with autologous tumor lysate-pulsed DC, an event associated with editing of TRP2 hi tumor cells and a heightened sensitivity of resampled tumors to chemotherapy agents in vitro including TMZ [17]. Survivin [18][19][20] and the mutant epidermal growth factor receptor EGFRvIII [21] are two other glioma-associated antigens also implicated in mediating resistance of GBM to standard treatments. Thus, while relative overexpression of certain proteins confers chemoresistance, this may also render cells more susceptible to antigen-specific immune attack. ...
There is no standard treatment for recurrent glioblastoma multiforme (GBM). Retreatment with temozolomide (TMZ) is one treatment option. We reasoned this could be more effective if combined with a vaccine that preferentially targeted TMZ-resistant cells. To test the feasibility and safety of such an approach, a phase 1 trial was conducted in which patients with GBM tumors relapsing after standard chemoradiotherapy were retreated with TMZ in combination with a vaccine consisting of monocyte-derived dendritic cells (DC) pulsed with autologous tumor cells that had previously been exposed to TMZ in vivo in the course of primary treatment. Of 14 participants, nine patients completed the initial phase of priming vaccinations and two cycles of TMZ, one proved to have radionecrosis, one rapidly progressed, and in three the yield of DC vaccine was insufficient to proceed with treatment. Other than expected toxicities related to TMZ, there were no adverse events attributable to the combined treatment. Two patients had objective radiological responses. Six month progression-free survival was 22 %, similar to retreatment with TMZ alone. Anti-tumor immune responses were assessed in peripheral blood mononuclear cells using interferon-γ ELISpot, with two patients meeting criteria for a vaccine-induced immune response, one of whom remained disease-free for nearly three years. Another patient with an anti-tumor immune response at baseline that was sustained post-vaccination experienced a 12-month period of progression-free survival. In summary, the combined treatment was safe and well-tolerated but feasibility in the recurrent setting was marginal. Evidence of immune responses in a few patients broadly correlated with better clinical outcome.
... Survivin is a member of the family of the inhibitor of apoptosis proteins and functions as a key regulator of mitosis and programmed cell death (24). Survivin is overexpressed in various types of tumors with the frequency of 34.5% in gastric cancers (25), 50-60% in colorectal cancers (25,26), 64% in malignant gliomas (27), 53-72% in lung cancers (28,29), and 70.7% in breast cancers (30). Cancer vaccines to induce an antigen-specific immune responses against survivin-expressing tumor cells have been developed with :01 eEF2 cells expression expression expression T2 --Undetectable T2-2402 + -Undetectable T2-0201 -+ Undetectable SW480 + + AZ-521 -+ MKN28 - ...
Recent studies have shown that cancer immunotherapy could be a promising therapeutic approach for the treatment of cancer. In the present study, to identify novel tumor-associated antigens (TAAs), the proteins expressed in a panel of cancer cells were serologically screened by immunoblot analysis and the eukaryotic elongation factor 2 (eEF2) was identified as an antigen that was recognized by IgG autoantibody in sera from a group of patients with head and neck squamous cell carcinoma (HNSCC) or colon cancer. Enzyme-linked immunosorbent assay showed that serum eEF2 IgG Ab levels were significantly higher in colorectal and gastric cancer patients compared to healthy individuals. Immunohistochemistry experiments showed that the eEF2 protein was overexpressed in the majority of lung, esophageal, pancreatic, breast and prostate cancers, HNSCC, glioblastoma multiforme and non-Hodgkin's lymphoma (NHL). Knockdown of eEF2 by short hairpin RNA (shRNA) significantly inhibited the growth in four eEF2-expressing cell lines, PC14 lung cancer, PCI6 pancreatic cancer, HT1080 fibrosarcoma and A172 glioblastoma cells, but not in eEF2-undetectable MCF7 cells. Furthermore, eEF2-derived 9-mer peptides, EF786 (eEF2 786-794 aa) and EF292 (eEF2 292-300 aa), elicited cytotoxic T lymphocyte (CTL) responses in peripheral blood mononuclear cells (PBMCs) from an HLA-A*24:02- and an HLA-A*02:01-positive healthy donor, respectively, in an HLA-A-restricted manner. These results indicated that the eEF2 gene is overexpressed in the majority of several types of cancers and plays an oncogenic role in cancer cell growth. Moreover, the eEF2 gene product is immunogenic and a promising target molecule of cancer immunotherapy for several types of cancers.
Gliomas are the most frequent type of tumor in the central nervous system, which exhibit properties that make their treatment difficult, such as cellular infiltration, heterogeneity, and the presence of stem-like cells responsible for tumor recurrence. The response of this type of tumor to chemoradiotherapy is poor, possibly due to a higher repair activity of the genetic material, among other causes. The DNA double-strand breaks are an important type of lesion to the genetic material, which have the potential to trigger processes of cell death or cause gene aberrations that could promote tumorigenesis. This review describes how the different cellular elements regulate the formation of DNA double-strand breaks and their repair in gliomas, discussing the therapeutic potential of the induction of this type of lesion and the suppression of its repair as a control mechanism of brain tumorigenesis.
Aim: Survivin is an "inhibitor of apoptosis" protein. Survivin expression is a poor prognostic factor in a variety of solid tumors. In this clinicopathological study, we aimed to investigate survivin immunostaining of leiomyomas, leiomyoma variants, STUMP (Uterine smooth muscle tumor of uncertain malignant potential)'s and LMS (leiomyosarcoma). Our second objective was to investigate whether survivin immunoreactivity in STUMP and LMS may play a role in determining recurrence. Material and Methods: Consecutive 119 specimens of leiomyoma, leiomyoma variants, STUMP and LMS from the pathology archives of Bezmialem Medical Faculty and Cerrahpasa Medical Faculty were selected. Clinicopathologic characteristics were analyzed and specimens were stained with survivin and Ki-67. The percentage and staining intensity of immunoreactive cells were examined. Additionally, we analyzed whether survivin intensity and expression might be a predictor of LMS recurrence. Results: The patients in the LMS group were older (p< 0.001). All LMS and all STUMP specimens were stained with survivin. Survivin staining and Ki-67 staining were highest in the LMS and STUMP groups. Survivin staining was 14.2 ± 6.7 % in the LMS group, 11.2± 10.4 % in the STUMP group, 1.85 ± 1.9 % in the leiomyoma group and 1.4 ± 0.2 % in the leiomyoma variant group (p<0.001). Survivin staining intensity was 1.2 ± 0.6 in the LMS group, 0.9 ± 0.2 in the STUMP group, 0.8 ± 0.4 in the leiomyoma group and 0.9 ± 0.3 in the leiomyoma variant group (p=0.025). Both survivin staining percentage and staining intensity correlated with the Ki-67 proliferation index. In the LMS cases that showed recurrence survivin staining was 16% while in the cases that did not reoccur survivin staining was 2% (p<0.001). Conclusion: The antiapoptotic marker "survivin" has not been studied before for smooth muscle tumors of the uterus. Utilizing survivin in conjuncture with histologic features and Ki-67 can also help to determine malignancy potential and LMS recurrence.
Immunotherapy applications to glioblastoma represent a new treatment frontier. Antigen-targeted immunotherapy approaches hold enormous potential to elicit antigen-specific anti-tumor effects in central nervous system tumors. Still, the paucity of effective antigen targets remains a significant obstacle in safely and effectively treating glioblastoma and other malignant gliomas with relatively low mutation loads. In this review, we highlight the current understanding of and development of immunotherapy to target 1) shared non-mutant antigens 2) shared mutant antigens (neoantigens) derived from cancer-specific mutations 3) personalized neoantigens derived from tumor-specific genetic alterations containing de novo peptide sequences and 4) virus-derived antigens. We also discuss strategies to enhance tumor immunogenicity and neoantigen prediction. Spatial heterogeneity remains a formidable challenge for immunotherapy of glioma; recent advances in targeting multiple antigens and refining the antigen selection pipeline hold great promise to turn the tide against glioma.
Glioblastoma (GBM) is one of the deadliest tumors and has a median survival of 3 months if left untreated. Despite advances in rationally targeted pharmacological approaches, the clinical care of GBM remains palliative in intent. Since the majority of altered signaling cascades involved in cancer establishment and progression eventually affect cell cycle progression, an alternative approach for cancer therapy is to develop innovative compounds that block the activity of crucial molecules needed by tumor cells to complete cell division. In this context, we review promising ongoing and future strategies for GBM therapeutics aimed towards G2/M inhibition such as anti-microtubule agents and targeted therapy against G2/M regulators like cyclin-dependent kinases, Aurora inhibitors, PLK1, BUB, 1, and BUBR1, and survivin. Moreover, we also include investigational agents in the preclinical and early clinical settings. Although several drugs were shown to be gliotoxic, most of them have not yet entered therapeutic trials. The use of either single exposure or a combination with novel compounds may lead to treatment alternatives for GBM patients in the near future.
Purpose:
Survivin is an inhibitor of apoptosis protein (IAP) that is highly expressed in many cancers and represents an attractive molecule for targeted cancer therapy. Although primarily regarded as an intracellular protein with diverse actions, survivin has also been identified in association with circulating tumor exosomes.
Experimental design:
We have reported that active specific vaccination with a long peptide surviving immunogen leads to the development of survivin-specific CD8-mediated tumor cell lysis and prolongation of survival in tumor-bearing mice. In addition to cellular antitumor responses, circulating anti-survivin antibodies are detected in the serum of mice and human glioblastoma patients following vaccination with the survivin immunogen.
Results:
Here we demonstrate that survivin is present on the outer cell membrane of a wide variety of cancer cell types, including both murine and human glioma cells. In addition, antibodies to surviving that are derived from the immunogen display antitumor activity against murine GL261 gliomas in both flank and intracranial tumor models and against B16 melanoma as well.
Conclusion:
In addition to immunogen-induced, CD8-mediated tumor cell lysis, antibodies to the survivin immunogen have antitumor activity in vivo. Cell-surface survivin could provide a specific target for antibody-mediated tumor immunotherapeutic approaches.
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.
The generation of a targeting agent that strictly binds to IL13Rα2 will significantly expand the therapeutic potential for the treatment of IL13Rα2-expressing cancers. In order to fulfill this goal, we generated a single-chain antibody (scFv47) from our parental IL13Rα2 monoclonal antibody and tested its binding properties. Furthermore, to demonstrate the potential therapeutic applicability of scFv47, we engineered an adenovirus by incorporating scFv47 as the targeting moiety in the viral fiber and characterized its properties in vitro and in vivo. The scFv47 binds to human recombinant IL13Rα2, but not to IL13Rα1 with a high affinity of 0.9 · 10(-9) M, similar to that of the parental antibody. Moreover, the scFv47 successfully redirects adenovirus to IL13Rα2 expressing glioma cells both in vitro and in vivo. Our data validate scFv47 as a highly selective IL13Rα2 targeting agent and justify further development of scFv47-modified oncolytic adenovirus and other therapeutics for the treatment of IL13Rα2-expressing glioma and other malignancies.
AIM: To investigate the expression of survivin in activated hepatic stellate cell (HSC) and the effect of transfection of antisense oligonucleotide (ASODN) targeting the survivin gene on apoptosis of HSC-T6 cells. METHODS: The experiment set the control group which not added liposome and oligonucleotide (ODN), liposome group which only added liposome, sense oligonucleotide (SODN) group (1 000 nmol/L) and 400, 800, 1 000 nmol/L ASODN group. The expression of survivin protein in activated HSC, hepatic carcinoma cells, and normal liver cells was detected by immunofluorescence. ASODN targeting the survivin gene was transfected into HSC-T6 cells with Lipofectamine™ 2000, and transfection efficiency was detected by fluorescence microscopy. Forty-eight hours after transfection, the changes in survivin mRNA and protein expression were assessed by RT-PCR and Western blot, respectively, and cell apoptosis was measured by PI staining and flow cytometry. RESULTS: The positive rate of survivin expression in activated HSC was significantly higher than that in normal liver cells (66.07% ± 8.55% vs 9.74% ± 2.68%, P < 0.05). The positive rate of survivin expression was highest in hepatic carcinoma cells among the three groups of cells (69.41% ± 9.10%). Strong green fluorescence was observed by fluorescence microscopy in cells transfected with different concentrations of ASODN, and the transfection efficiency reached 80%. Compared with blank control cells and cells transfected with empty lipofectamine™ 2000 or control oligonucleotide, the expression levels of survivin mRNA (0.94 ± 0.03, 0.95 ± 0.04 and 0.92 ± 0.04 vs 0.64 ± 0.02, 0.54 ± 0.02 and 0.26 ± 0.01) and protein (0.84 ± 0.02, 0.82 ± 0.03 and 0.81 ± 0.02 vs 0.53 ± 0.02, 0.38 ± 0.01 and 0.20 ± 0.01) were significantly decreased (all P < 0.01), and the apoptosis rate (19.00% ± 0.53%, 29.80% ± 1.54% and 48.70% ± 2.00%, respectively) significantly increased (all P < 0.01) in cells transfected with different concentrations of ASODN. CONCLUSION: Survivin is highly expressed in activated HSC-T6 cells. Down-regulation of survivin expression by transfection of ASODN targeting the survivin gene can induce apoptosis of HSC-T6 cells.
Over the past two decades, small interfering RNA (siRNA) has gained widespread popularity as an efficient tool for cancer therapy. Unfortunately, due to its inherent instability, the progress in the field has yet to hit a benchmark with no marketed siRNA formulations for cancer, yet. siRNA delivery via nonviral carriers have recently generated immense interest and this chapter is an attempt at a comprehensive coverage of liposomes and micelles as non-viral siRNA delivery systems. The myriad of options and recent advancements with respect to these nanocarriers in cancer therapy are discussed here.
Most anticancer therapies commonly used in the treatment of human cancer, e.g., chemotherapy, γ-irradiation, immunotherapy, or suicide gene therapy, kill tumor cells by triggering cell death pathways including apoptosis in cancer cells. Hence, the failure to activate such pathways may lead to the resistance of cancers to current treatment approaches. A better understanding of the molecular events that regulate apoptosis in cancers and upon cancer therapy provides the basis for a more rational approach to developing molecular targeted therapies in the fight against cancer.
A survivin small interfering RNA sequence specific for a human and mouse homogenous sequence was constructed. Survivin small interfering RNA could significantly inhibit glioma cell proliferation and induce apoptosis when it was transfected into either a human glioma cell line U251 or rat glioma C6 cells in vitro. In addition, treatment of rat orthotopic glioma models with survivin small interfering demonstrated the inhibition of glioma growth in vivo. Our experimental findings suggest that the use of RNA interference techniques to target the survivin sequence may be useful in the treatment of glioma.
Glioblastoma multiforme (GBM) has a poor prognosis despite maximal multimodal therapy. Biomarkers of relevance to prognosis which may also identify treatment targets are needed. A few hundred genetic and molecular predictors have been implicated in the literature, however with the exception of IDH1 and O6-MGMT, there is uncertainty regarding their true prognostic relevance. This study analyses reported genetic and molecular predictors of prognosis in GBM. For each, its relationship with univariate overall survival in adults with GBM is described. A systematic search of MEDLINE (1998-July 2010) was performed. Eligible papers studied the effect of any genetic or molecular marker on univariate overall survival in adult patients with histologically diagnosed GBM. Primary outcomes were median survival difference in months and univariate hazard ratios. Analyses included converting 126 Kaplan-Meier curves and 27 raw data sets into primary outcomes. Seventy-four random effects meta-analyses were performed on 39 unique genetic or molecular factors. Objective criteria were designed to classify factors into the categories of clearly prognostic, weakly prognostic, non-prognostic and promising. Included were 304 publications and 174 studies involving 14,678 unique patients from 33 countries. We identified 422 reported genetic and molecular predictors, of which 52 had ⩾2 studies. IDH1 mutation and O6-MGMT were classified as clearly prognostic, validating the methodology. High Ki-67/MIB-1 and loss of heterozygosity of chromosome 10/10q were classified as weakly prognostic. Four factors were classified as non-prognostic and 13 factors were classified as promising and worthy of additional investigation. Funnel plot analysis did not identify any evidence of publication bias. This study demonstrates a novel literature and meta-analytical based approach to maximise the value that can be derived from the plethora of literature reports of molecular and genetic factors in GBM. Caution is advised in over-interpreting the results due to study limitations. Further research to develop this methodology and improvements in study reporting are suggested.
Copyright © 2014 Elsevier Ltd. All rights reserved.
Survivin has multiple functions in the progression of cancer. The aim of the present study was to investigate the influence of YM155, a specific survivin inhibitor, on the biological behavior of U87 glioblastoma cells. The proliferative activity and growth rate of U87 cells were determined by colony formation assay and mononuclear cell direct cytotoxicity assay (MTT assay). The reconstituted basement membrane penetrating capacity was determined by cell invasion assay. The cell movement and migratory capacity were detected by wound-healing repair assay. We found that inhibition of survivin by YM155 dramatically decreased the invasive and metastatic capacities of the cells, suppressed the proliferation, decelerated the rate of growth, and reduced the number of clones on soft agar. In conclusion, YM155 has the potential to be used in the clinical treatment of GBM.
Survival for patients with glioblastoma, the most common high-grade primary CNS tumor, remains poor despite multiple therapeutic
interventions including intensifying cytotoxic therapy, targeting dysregulated cell signaling pathways, and blocking angiogenesis.
Exciting, durable clinical benefits have recently been demonstrated for a number of other challenging cancers using a variety
of immunotherapeutic approaches. Much modern research confirms that the CNS is immunoactive rather than immunoprivileged.
Preliminary results of clinical studies demonstrate that varied vaccine strategies have achieved encouraging evidence of clinical
benefit for glioblastoma patients, although multiple variables will likely require systematic investigation before optimal
outcomes are realized. Initial preclinical studies have also revealed promising results with other immunotherapies including
cell-based approaches and immune checkpoint blockade. Clinical studies to evaluate a wide array of immune therapies for malignant
glioma patients are being rapidly developed. Important considerations going forward include optimizing response assessment
and identifiying correlative biomarkers for predict therapeutic benefit. Finally, the potential of complementary combinatorial
immunotherapeutic regimens is highly exciting and warrants expedited investigation.
Up to now, survivin has been recommended as a prognostic and diagnostic indicator in glioma patients. However, there are still many controversies. Here, a meta-analysis was conducted to draw a more definitive conclusion on the correlation of survivin with overall survival (OS), age, gender, and WHO grade. Eligible studies were available through careful assessment, and then pooled hazard ratios (HRs) or odds ratios (ORs) with 95 % confidence intervals (95 % CIs) were estimated. Funnel plots were introduced to evaluate the publication bias. Additionally, heterogeneity and sensitivity were also evaluated. In the present meta-analysis, 15 eligible studies with a total of 1,089 patients were incorporated. Survivin expression in gliomas correlated with 2-year OS (n = 8; HR 0.17, 95 % CI 0.11-0.26) and 5-year OS (n = 7; HR 0.12, 95 % CI 0.07-0.22) in patients. In addition, a fixed-effect model revealed a significant association between survivin and age (male/+; OR 2.10, 95 % CI 1.44-3.05) and survivin and WHO grade (I+II/+; OR 0.27, 95 % CI 0.19-0.38). No heterogeneity was observed across all studies. According to Begg's and Egger's test and funnel plot, no publication bias was reported. Taken together, our meta-analysis suggests that survivin expression is associated with poor survival, older age, and higher WHO grade and could be suggested as a useful prognostic and diagnostic biomarker, or an effective therapy target.
The majority of human malignant glioma cells express Fas/APO-1 and are susceptible to Fas/APO-1 antibody-mediated apoptosis in vitro. The sensitivity of Fas/APO-1-positive glioma cell lines to Fas/APO-1 antibody-mediated killing correlates inversely with the constitutive expression of the antiapoptotic protooncogene bcl-2. Here we report that BCL-2 protein expression of human glial tumors in vivo correlates with malignant transformation in that BCL-2 immunoreactive glioma cells were more abundant in WHO grade III/IV gliomas than in grade I/II gliomas. Fas/APO-1 antibody-sensitive human glioma cell lines stably transfected with a murine bcl-2 cDNA acquired resistance to Fas/APO-1 antibody-mediated apoptosis. Forced expression of bcl-2 also attenuated TNF alpha-mediated cytotoxicity of glioma cell lines in the presence of actinomycin D and cycloheximide and conferred partial protection from irradiation and the cancer chemotherapy drugs, cisplatin and BCNU. Preexposure of the glioma cell lines to the cytokines, IFN gamma and TNF alpha, which sensitize for Fas/APO-1-dependent killing, partially overcame bcl-2-mediated rescue from apoptosis, suggesting that multimodality immunotherapy involving cytokines and Fas/APO-1 targeting might eventually provide a promising approach to the treatment of human malignant gliomas.
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.
Survivin is a new IAP apoptosis inhibitor expressed during development and in human cancer in vivo. The coding strand of the survivin gene was extensively complementary to that of effector cell protease receptor-1 (EPR-1), prompting the present investigation
on the origin and functional relationship of these two transcripts. Southern blots of genomic DNA were consistent with the
presence of multiple, evolutionarily conserved, EPR-1/Survivin-related genes. By pulsed field gel electrophoresis and single-
and two-color fluorescence in situ hybridization, these were contained within a contiguous physical interval of 75–130 kilobases (kb) on chromosome 17q25. In
Northern blots, a single strand-specific probe identified a 1.3-kb EPR-1 mRNA broadly distributed in normal adult and fetal
tissues, structurally distinct from the 1.9-kb Survivin transcript expressed in transformed cell lines. Transient co-transfection
of an EPR-1 cDNA potentially acting as a Survivin antisense with a lacZ reporter plasmid resulted in loss of viability of HeLa cells. In contrast, co-transfection of an antisense cDNA of intercellular
adhesion molecule-1 or a sense-oriented Survivin cDNA was without effect. In stably transfected HeLa cells, ZnSO4 induction of an EPR-1 mRNA under the control of a metallothionein promoter suppressed the expression of endogenous survivin.
This resulted in (i) increased apoptosis as detected by analysis of DNA content and in situ internucleosomal DNA fragmentation and (ii) inhibition of cell proliferation as compared with induced vector control transfectants.
These findings suggest the existence of a potential EPR-1/survivin gene cluster and identify survivin as a new target for disrupting cell viability pathways in cancer.
Background/methods:
Gliomas are common malignant neoplasms of the central nervous system. Among the major subtypes of gliomas, oligodendrogliomas are distinguished by their remarkable sensitivity to chemotherapy, with approximately two thirds of anaplastic (malignant) oligodendrogliomas responding dramatically to combination treatment with procarbazine, lomustine, and vincristine (termed PCV). Unfortunately, no clinical or pathologic feature of these tumors allows accurate prediction of their response to chemotherapy. Anaplastic oligodendrogliomas also are distinguished by a unique constellation of molecular genetic alterations, including coincident loss of chromosomal arms 1p and 19q in 50%-70% of tumors. We have hypothesized that these or other specific genetic changes might predict the response to chemotherapy and prognosis in patients with anaplastic oligodendrogliomas. Therefore, we have analyzed molecular genetic alterations involving chromosomes 1p, 10q, and 19q and the TP53 (on chromosome 17p) and CDKN2A (on chromosome 9p) genes, in addition to clinicopathologic features in 39 patients with anaplastic oligodendrogliomas for whom chemotherapeutic response and survival could be assessed.
Results/conclusions:
Allelic loss (or loss of heterozygosity) of chromosome 1p is a statistically significant predictor of chemosensitivity, and combined loss involving chromosomes 1p and 19q is statistically significantly associated with both chemosensitivity and longer recurrence-free survival after chemotherapy. Moreover, in both univariate and multivariate analyses, losses involving both chromosomes 1p and 19q were strongly associated with longer overall survival, whereas CDKN2A gene deletions and ring enhancement (i.e., contrast enhancement forming a rim around the tumor) on neuroimaging were associated with a significantly worse prognosis. The inverse relationship between CDKN2A gene deletions and losses of chromosomes 1p and 19q further implies that these differential clinical behaviors reflect two independent genetic subtypes of anaplastic oligodendroglioma. These results suggest that molecular genetic analysis may aid therapeutic decisions and predict outcome in patients with anaplastic oligodendrogliomas.
IAP family proteins are conserved throughout animal evolution and can block apoptosis when expressed in cells derived from multiple species. In many instances IAP family proteins can suppress apoptosis across species barriers (for review, see Clem and Duckett 1998), implying that although the details of their regulation may vary, these proteins evidently target a common mechanism involved in programmed cell death. Although all IAP family proteins require at least one BIR domain for their anti-apoptotic function, it should be emphasized that not all BIR-containing proteins are necessarily involved in apoptosis regulation as indicated by the failure of the Ac-IAP protein to suppress apoptosis despite harboring a BIR domain. Until proven otherwise, the most likely explanation for how IAPs prevent apoptosis is by binding to and inhibiting caspases, as indicated by recent studies (Deveraux et al. 1997 1998; Roy et al. 1997; Takahashi et al. 1998; Tamm et al. 1998). Assuming that this biased viewpoint is correct, how important are IAP family proteins likely to be for ensuring cell survival in vivo? The answer to that question probably depends on the type of cell under investigation and the specific cell death stimulus involved. I IAPs function primarily as inhibitors of caspases, then we can anticipate from other experiments where artificial means were used to suppress these proteases that APs will be capable of rescuing cells from some cell death signals but not others (Green and Reed 1998). Mitochondrial involvement appears to be one of the key variables that determines whether caspase inhibitors are sufficient to provide long-term protection and preservation of clonigenic potential, versus merely delaying death by converting an apoptotic stimulus into a necrotic one (Reed 1997; Green and Reed 1998). In many types of cells, loss of cytochrome c from mitochondria, for example, has two ways of killing cells: (1) activation of caspases through Apaf-1; or (2) cessation of mitochondrial electron chain transport with subsequent ATP depletion, generation of reactive oxygen species, and related sequelae. If the role of IAPs is relegated to caspase suppression, this may prevent cytochrome c-induced apoptosis, but not necessarily stop cell death induced by caspase-independent mechanisms. Recent studies in which release of cytochrome c was found to be a potentially reversible event suggest that whether cytochrome c loss from mitochondria defines a cell death commitment point will likely vary among cell types and depending on a variety environmental factors (Q. Chen et al. 1998). These factors may include the extent to which cells are able to produce sufficient ATP from anaerobic glycolysis in the cytosol and the method by which mitochondrial membrane barrier function was altered to allow for exodus of cytochrome c (i.e., reversible versus irreversible/rupture). Defining the in vivo requirements for IAPs in the maintenance of cell survival may be difficult because of potential redundancy. Humans have at least five and possibly more IAP family genes and even lower organisms, such as Drosophila, appear to contain at least two IAP genes, implying evolutionary pressure to ensure adequate back-up if one of these genes were to become inactivated. If IAPs do indeed function predominantly as caspase inhibitors, then one could imagine a very important role for these endogenous protease inhibitors in ensuring that the small amounts of adventitial caspase activation, which must surely occur on a routine basis, do not amplify out of control, resulting in inappropriate cell death. In this regard, virtually every other protease system studied to date contains molecules whose sole function is directed toward dampening proteolysis through the cascade (for review, see Colman et al. 1994), thus ensuring that biologically appropriate triggering of the pathway only occurs when certain thresholds are surpassed. By analogy, it is attractive to consider IAP family proteins in the same way, as proteins that set thresholds for how much caspase activation is necessary to successfully trigger apoptosis. Through alterations in the levels of IAP family gene expression and interactions of IAPs with other proteins, this IAP-dependent threshold for caspase-induced apoptosis could be varied to suit various physiological needs. Dysregulation of these normal control mechanisms then could be a contributor to various diseases characterized by excessive (ischemia, AIDS, SMA) or inadequate (cancer, autoimmunity) cell death.
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.
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.
In many human cancers, the INK4A locus is frequently mutated by homozygous deletions. By alternative splicing this locus encodes two non-related tumor suppressor genes, p16(INK4A) and p14(ARF) (p19(ARF) in mice), which regulate cell cycle and cell survival in the retinoblastoma protein (pRb) and p53 pathways, respectively. In mice, the role of p16(INK4A) as the critical tumor suppressor gene at the INK4A locus was challenged when it was found that p19(ARF) only knock-out mice developed tumors, including gliomas. We have analysed the genetic status of the INK4A locus in 105 primary gliomas using both microsatellite mapping (MSM) and quantitative real-time PCR (QRT-PCR). Comparison of the results of the two methods revealed agreement in 67% of the tumors examined. In discordant cases, fluorescence in situ hybridization (FISH) analysis was always found to support QRT-PCR classification. Direct assessment of p14(ARF) exon 1beta, p16(INK4A) exon 1alpha and exon 2 by QRT-PCR revealed 43 (41%) homozygous and eight (7%) hemizygous deletions at the INK4A locus. In 49 (47%) gliomas, both alleles were retained. In addition, QRT-PCR, but not MSM, detected hyperploidy in five (5%) tumors. Deletion of p14(ARF) was always associated with co-deletion of p16(INK4A) and increased in frequency upon progression from low to high grade gliomas. Shorter survival was associated with homozygous deletions of INK4A in the subgroup of glioblastoma patients older than 50 years of age (P=0.025, Anova test single factor, alpha=0.05).
Malignant brain tumors strike deep into the psyche of those receiving and those delivering the diagnosis. Ma- lignant gliomas, the most common subtype of primary brain tumors, are aggressive, highly invasive, and neuro- logically destructive tumors considered to be among the deadliest of human cancers. In its most aggressive mani- festation, glioblastoma (GBM), median survival ranges from 9 to 12 months, despite maximum treatment ef- forts—a statistical fact that has changed little over sev- eral decades of technological advances in neurosurgery, radiation therapy, and clinical trials of conventional and novel therapeutics. Over the same time period, there has been an explosion of knowledge in cancer biology and basic science discovery that has fueled meaningful prog- ress in the treatment of many common human cancers, including those of the breast, lung, and prostate. It is perplexing that therapies used effectively in the treat- ment of these solid tumors are overwhelmingly ineffec- tive in the treatment of GBM, perhaps reflecting the ec- centric biology and cellular origin of this neoplasm. To date, only one new agent has been documented to have modest activity against intermediate-grade gliomas, whereas no effective agents have emerged for the treat- ment of GBM, despite 20 years of enrolling patients in clinical trials. It is ironic that although a comprehensive view of the genetic lesions encountered in malignant gliomas has been compiled, substantive conceptual and practical barriers remain in assigning functional signifi- cance to these genetic changes and in harnessing this basic information into the development of drugs that make a difference in patient care. The history of treating malignant gliomas dates back to the middle of the 19th century and parallels landmark advances in modern surgical technique and the clinical discipline of neurology. The first brain tumor surgery of the modern era was performed in 1884 by Rickman
A controlled, prospective, randomized study evaluated the use of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and/or radiotherapy in the treatment of patients who were operated on and had histological confirmation of anaplastic glioma. A total of 303 patients were randomized into this study, of whom 222 (73%) were within the Valid Study Group (VSG), having met the protocol criteria of neuropathology, corticosteroid control, and therapeutic approach. Patients were divided into four random groups, and received BCNU (80 mg/sq m/day on 3 successive days every 6 to 8 weeks), and/or radiotherapy (5000 to 6000 rads to the whole brain through bilateral opposing ports), or best conventional care but no chemotherapy or radiotherapy. Analysis was performed on all patients who received any amount of therapy (VSG) and on the Adequately Treated Group (ATG), who had received 5000 or more rads radiotherapy, two or more courses of chemotherapy, and had a minimum survival of 8 or more weeks (the interval that would have been required to have received either the radiotherapy or chemotherapy). Median survival of patients in the VSG was, best conventional care: 14 weeks (ATG: 17.0 weeks); BCNU: 18.5 weeks (ATG: 25.0 weeks); radiotherapy: 35 weeks (ATG: 37.5 weeks); and BCNU plus radiotherapy: 34.5 weeks (ATG: 40.5 weeks). All therapeutic modalities showed some statistical superiority compared to best conventional care. There was no significant difference between the four groups in relation to age distribution, sex, location of tumor, diagnosis, tumor characteristics, signs or symptoms, or the amount of corticosteroid used. An analysis of prognostic factors indicates that the initial performance status (Karnofsky rating), age, the use of only a surgical biopsy, parietal location, the presence of seizures, or the involvement of cranial nerves II, III, IV, and VI are all of significance. Toxicity included acceptable, reversible thrombocytopenia and leukopenia.
Within three weeks of definitive surgical intervention, 467 patients with histologically proved malignant glioma were randomized to receive one of four treatment regimens: semustine (MeCCNU), radiotherapy, carmustine (BCNU) plus radiotherapy, or semustine plus radiotherapy. We analyzed the data for the total randomized population and for the 358 patients in whom the initial protocol specifications were met (the valid study group). Observed toxicity included acceptable skin reactions secondary to radiotherapy and reversible leukopenia and thrombocytopenia due to chemotherapy. Radiotherapy used alone or in combination with a nitrosourea significantly improved survival in comparison with semustine alone. The group receiving carmustine plus radiotherapy had the best survival, but the difference in survival between the groups receiving carmustine plus radiotherapy and semustine plus radiotherapy was not statistically significant. The combination of carmustine plus radiotherapy produced a modest benefit in long-term (18-month) survival as compared with radiotherapy alone, although the difference between survival curves was not significiant at the 0.05 level. This study suggests that it is best to use radiotherapy in the post-surgical treatment of malignant glioma and to continue the search for an effective chemotherapeutic regimen to use in addition to radiotherapy.
The Bax protein plays a critical role in the apoptosis of cancers induced by radiotherapy or chemotherapy, which induce both apoptosis and necrosis. We transduced various glioblastoma cells with the Bax gene via an adenoviral vector and found that A-172 cells led to necrotic cell death, while U251 cells apoptotic cell death, even though a similar level of Bax protein was introduced. A-172 cells displayed a much higher constitutive expression of the Bcl-XL protein compared with that of U251 cells. Upon simultaneous overexpression of the Bcl-XL and Bax proteins in the U251 cells, Bax-induced apoptosis of U251 cells was suppressed and an increase in the number of necrotic cells was seen. Moreover, induction of a higher amount of Bax protein in A-172 cells increased the percentage of apoptotic cells. In conclusion, if a cancerous cell expresses a high enough amount of Bax to undergo death, apoptosis will be induced. If a cancerous cell expresses a level of Bcl-XL which prevents Bax-induced apoptosis, the overexpression of Bax leads to necrotic cell death.
Survivin is observed uniquely in tumor cells and developmental cells, which undergo either inappropriate or programmed cell growth. In the current study, we investigated the influence of Survivin on cell cycle. Overexpression of Survivin resulted in accelerated S phase shift, resistance to G1 arrest, and activated Cdk2/Cyclin E complex leading Rb phosphorylation. In addition, nuclear translocation of Survivin followed by an interaction with Cdk4 was detected. Interestingly, Survivin nuclear translocation coincided with S phase shift, and prevention of nuclear transport suppressed Survivin nuclear translocation and S phase shift. Further, we also observed that Survivin competitively interacted with the Cdk4/p16(INK4a) complex in a cell free system and in vivo. These results suggest that Survivin initiates the cell cycle entry as a result of nuclear translocation followed by an interaction with Cdk4.
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.
Dysregulation of apoptosis may favor onset and progression of cancer and influence response to therapy. Survivin is an inhibitor of apoptosis that is selectively overexpressed in common human cancers, but not in normal tissues, and that correlates with aggressive disease and unfavorable outcomes.
To investigate the potential suitability of survivin detection in urine as a novel predictive/prognostic molecular marker of bladder cancer.
Survey of urine specimens from 5 groups: healthy volunteers (n = 17) and patients with nonneoplastic urinary tract disease (n = 30), genitourinary cancer (n = 30), new-onset or recurrent bladder cancer (n = 46), or treated bladder cancer (n = 35), recruited from 2 New England urology clinics.
Detectable survivin levels, analyzed by a novel detection system and confirmed by Western blot and reverse transcriptase polymerase chain reaction (RT-PCR), in urine samples of the 5 participant groups.
Survivin was detected in the urine samples of all 46 patients with new or recurrent bladder cancer using a novel detection system (31 of 31) and RT-PCR (15 of 15) methods. Survivin was not detected in the urine samples of 32 of 35 patients treated for bladder cancer and having negative cystoscopy results. None of the healthy volunteers or patients with prostate, kidney, vaginal, or cervical cancer had detectable survivin in urine samples. Of the 30 patients with nonneoplastic urinary tract disease, survivin was detected in 3 patients who had bladder abnormalities noted using cystoscopy and in 1 patient with an increased prostate-specific antigen level. Patients with low-grade bladder cancer had significantly lower urine survivin levels than patients with carcinoma in situ (P =.002).
Highly sensitive and specific determination of urine survivin appears to provide a simple, noninvasive diagnostic test to identify patients with new or recurrent bladder cancer.
Beneath the complexity and idiopathy of every cancer lies a limited number of 'mission critical' events that have propelled the tumour cell and its progeny into uncontrolled expansion and invasion. One of these is deregulated cell proliferation, which, together with the obligate compensatory suppression of apoptosis needed to support it, provides a minimal 'platform' necessary to support further neoplastic progression. Adroit targeting of these critical events should have potent and specific therapeutic consequences.