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SDS-PAGE banding patterns of TSP-180 from J:PO4 labeled control Line I cells (lanes I, 2) or Line I cells after the addition of 0.2 /ig/ml of MoAb I35-13C (lanes 3. 4) to the culture medium. Lanes I, 3: MoAb 135-13C: Lanes 2. 4: rabbit antiserum to rat IgG. Arrows as indicated in Fig. 1.
Source publication
A tumor surface protein (TSP-180) has been identified on murine lung carcinomas using two monoclonal antibodies (MoAbs) (135-13C and 346-11A). Quantitative analysis of TSP-180 on 3LL variants maintained either in vitro or in vivo indicates that TSP-180 is highly expressed in highly malignant metastatic cells. In reducing conditions, sodium dodecyl...
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Citations
... Few studies have explored the role of integrin α6 in lung cancer, especially lung adenocarcinoma. The literature has reported that integrin α6β4 is up-regulated in murine Lewis lung carcinoma variants with high metastasis capacity (Sacchi et al. 1989). Jandova et al.'s (2015) study on various NSCLC cell lines has shown that up-regulation of integrin α6 is associated with elevated invasive potential and metastatic capacity of non-small lung adenocarcinoma cells. ...
... Earlier studies have revealed an association between integrin α6 and metastasis capacity in lung cancer. It has been reported that integrin α6β4 is up-regulated in murine Lewis lung carcinoma variants with high metastasis capacity (Sacchi et al. 1989). In addition, up-regulation of integrin α6 was shown to be related to the promotion of metastasis in lung cancer or non-small lung adenocarcinoma cell line (Hsu et al. 2013). ...
Objective
To study integrin α6 expression in lung adenocarcinoma tissue through comparison with matching adjacent non-cancerous tissues as well as elucidating the correlation between integrin α6 expression with the clinical parameters of lung adenocarcinoma. We also explore the signal pathways associated with integrin α6 up-regulation.
Methods
The clinical data, cancer tissues, and adjacent non-cancerous tissues of 30 patients diagnosed with lung adenocarcinoma were collected from Taizhou Hospital in Zhejiang Province, China, in 2010. The protein levels of integrin α6 were determined by immunohistochemistry methods. mRNA data of 85 lung adenocarcinoma tissues and 14 normal tissues as well as clinical results were collected from GEO30219. We also collected mRNA data of 533 lung adenocarcinoma tissues and 59 normal tissues as well as the clinical results of 522 patients with lung adenocarcinoma from the Cancer Genome Atlas (TCGA) database. The differences in protein and mRNA levels in cancer tissues and non-cancerous tissues were analyzed, and we subsequently investigated the association between integrin α6 expression and key parameters indicating lung adenocarcinoma progression and overall survival rate. Additionally, the possible pathways involved in the up-regulation of integrin α6 were analyzed by GSEA.
Results
The protein levels of integrin α6 in lung adenocarcinoma tissues were significantly higher than those in adjacent tissues (p < 0.01), and were positively correlated with the grade and T stage of lung adenocarcinoma (p < 0.05). Patients with low integrin α6 protein levels had higher survival rates (p < 0.05). The analysis of gene chip data from the TCGA database also showed that the integrin α6 mRNA level was significantly correlated with T stage (p < 0.05), overall survival (OS) rate (p < 0.01), and disease-free survival (DFS) rate (p = 0.005). GSEA gene enrichment analysis identified a series of pathways that may be associated with integrin α6 up-regulation, including the AGR, PYK2, ECM, and PTEN pathways.
Conclusion
Integrin α6 plays an important role in the occurrence and progression of lung adenocarcinoma and may act as a prognostic predictor of lung adenocarcinoma in patients. Based on the results of the present study, integrin α6 may be a potential target gene for the treatment of lung adenocarcinoma.
... The β4 integrin (ITGB4) subunit was initially termed as a tumour-associated antigen (TSP180) associated with metastasis [18,19]. The ITGB4 subunit binds exclusively to the α6 integrin subunit, forming a heterodimer of α6β4 integrin [20]. ...
Tumour budding predicts survival of stage II colorectal cancer (CRC) and has been suggested to be associated with epithelial‐to‐mesenchymal transition (EMT). However, the underlying molecular changes of tumour budding remain poorly understood. Here, we performed multiplex immunohistochemistry (mIHC) to phenotypically profile tumours using known EMT‐associated markers: E‐cadherin (adherence junctions), integrin β4 (ITGB4; basement membrane), ZO‐1 (tight junctions), and pan‐cytokeratin. A subpopulation of patients showed high ITGB4 expression in tumour buds, and this coincided with a switch of ITGB4 localisation from basal membrane of intact epithelium to the cytoplasm of budding cells. Digital image analysis revealed that tumour budding with high ITGB4 expression in tissue microarray (TMA) cores correlated with tumour budding assessed from H&E whole‐sections and independently predicted poor disease‐specific survival in two independent stage II CRC cohorts (hazard ratio (HR) =4 .50 (95% CI=1.50–13.5), n=232; HR=3.52 (95% CI=1.30‐9.53), n=72). Furthermore, digitally obtained ITGB4‐high bud count in random TMA cores associated better with survival outcome than visual tumour bud count in corresponding H&E stained samples. In summary, the mIHC‐based phenotypic profiling of human tumour tissue shows strong potential for the molecular characterisation of tumour biology and for the discovery of novel prognostic biomarkers.
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... The PI3K activation response to integrin α6β4 ligation is involved in invasive potential of carcinoma cells, and Tyr 1494 in the cytoplasmic domain of the integrin β4 is required for the activation [33]. Ligand binding to the extracellular domain of integrin α6β4 induced phosphorylation at serine and tyrosine residues in integrin β4 cytoplasmic domain, which were associated with a metastatic phenotype of cancer cells [34]. Phosphorylation of Tyr 1494 and Tyr 1526 in integrin β4 leads to recruitment of tyrosine phosphatase Shp2 and Shc to the β4 cytoplasmic domain, respectively, followed by activation of Ras-MAP kinase pathways, and promotes cell cycle progression [31,[35][36][37] (Figure 1). ...
Malignant transformation is accompanied with aberrant glycosylation of proteins. Such changes in glycan structure also occur in the integrins, which are a large family of cell surface receptors for the extracellular matrix and play key roles in tumor progression. There is now increasing evidence that glycosylation of integrins affects cellular signaling and interaction with the extracellular matrix, receptor tyrosine kinases, and galectins, thereby regulating cell adhesion, motility, growth, and survival. Integrin α6β4 is a receptor for laminin-332 and the increased expression level is correlated with malignant progression and poor survival in various types of cancers. Recent studies have revealed that integrin α6β4 plays central roles in tumorigenesis and the metastatic process. In this review, we summarize our current understanding of the molecular mechanisms of tumor progression driven by integrin α6β4 and also discuss the modification of glycans on integrin β4 subunit to address the important roles of glycan in integrin-mediated tumor progression.
... Our data support previous studies regarding integrin α6β4 and lung cancer progression. Early reports demonstrated that expression of the integrin α6β4 (previously described as TSP-180) is elevated in murine Lewis lung carcinoma variants with high metastatic potential [25]. Multiple studies investigating integrin β4 expression in patient-derived tissues have shown that it is expressed in NSCLC, with high levels observed in SCC [13][14][15]. ...
Lung cancer carries a poor prognosis and is the most common cause of cancer-related death worldwide. The integrin α6β4, a laminin receptor, promotes carcinoma progression in part by cooperating with various growth factor receptors to facilitate invasion and metastasis. In carcinoma cells with mutant TP53, the integrin α6β4 promotes cell survival. TP53 mutations and integrin α6β4 overexpression co-occur in many aggressive malignancies. Due to the high frequency of TP53 mutations in lung squamous cell carcinoma (SCC), we sought to investigate the association of integrin β4 expression with clinicopathologic features and survival in non-small cell lung cancer (NSCLC). We constructed a lung cancer tissue microarray and stained sections for integrin β4 subunit expression using immunohistochemistry. We found that integrin β4 expression is elevated in SCC compared to adenocarcinoma (P<.0001), which was confirmed in external gene expression datasets (P<.0001). We also determined that integrin β4 overexpression associates with the presence of venous invasion (P=.0048), and with reduced overall patient survival (Hazard ratio 1.46, 95% confidence interval 1.01 to 2.09, P=.0422). Elevated integrin β4 expression was also shown to associate with reduced overall survival in lung cancer gene expression datasets (Hazard ratio 1.49, 95% confidence interval 1.31 to 1.69, P<.0001). Using cBioPortal, we generated a network map demonstrating the 50 most highly altered genes neighboring ITGB4 in SCC which included laminins, collagens, CD151, genes in the EGFR and PI3K pathways, and other known signaling partners. In conclusion, we demonstrate that integrin β4 is overexpressed in NSCLC where it is an adverse prognostic marker.
... The upregulated integrin b1 level contributes to carcinogenesis and EMT progression [11,[13][14][15]. Unlike b1, integrin b4 displays completely different functions, either proadhesion or pro-invasiveness, which depends on tumor cell types [16,17]. The alteration of integrin b4 level is also linked with the initiation of EMT [4,18]. ...
Integrins have been known to play pivotal roles in malignant progression and epithelial-mesenchymal transition (EMT) of non-small cell lung cancer (NSCLC). We previously demonstrated that MARVELD1, a potential tumor suppressor, is epigenetically silenced in multiple cancer cells. In this study, we found MARVELD1 silencing altered cell surface ultrastructure of NSCLC cells and inhibited the formation of punctate integrin β1/β4 cluster in microvillus, whereas MARVELD1 overexpression suppressed TGF-β1-induced EMT. Remarkably, the balance of integrin β1 and β4 was modulated by MARVELD1. MARVELD1 silencing led to imbalance of integrin β1/β4 and significantly reduced microvillus length, furthermore affected the localization of β1/β4 at microvilli tips. TGF-β1-induced EMT was promoted by MARVELD1 silencing, while rebalance of integrin β1/β4 partly rescued the epithelial phenotype of MARVELD1-silenced cells. Mechanistically, we demonstrate that MARVELD1-mediated balance of integrin β1 and β4 regulates cell surface ultrastructure and EMT phenotype of NSCLC cells, suggesting MARVELD1 has a potential to be developed as a therapeutic target for NSCLC. © 2015 Wiley Periodicals, Inc.
... The therapeutic efficacy of chemotherapy and PDT can be greatly improved by efficient delivery of the drugs to the specific tumor location. The recent molecularly-targeting approach allows the medical intervention to affect only cancer cells but not the normal cells, based on molecular recognition processes (ligand-receptor or antibody-antigene interaction) [27][28][29][30][31]. This innovative approach is inherently different from classical modalities. ...
Treatment of brain cancer remains a challenge despite recent improvements in surgery and multimodal adjuvant therapy. Drug therapies of brain cancer have been particularly inefficient, due to the blood-brain barrier and the non-specificity of the potentially toxic drugs. The nanoparticle has emerged as a potential vector for brain delivery, able to overcome the problems of current strategies. Moreover, multi-functionality can be engineered into a single nanoplatform so that it can provide tumor-specific detection, treatment, and follow-up monitoring. Such multitasking is not possible with conventional technologies. This review describes recent advances in nanoparticle-based detection and therapy of brain cancer. The advantages of nanoparticle-based delivery and the types of nanoparticle systems under investigation are described, as well as their applications.
... Loss of Tumorigenicity-Since the tumorigenic and metastatic potentials of tumors are greatly affected by the expression levels of integrins (17)(18)(19)(20), we were interested in the behavior of the SM3-expressing cells in vivo. When the mock and the CST transfectants were inoculated subcutaneously into syngeneic C57/BL6 mice, and then examined for their tumor growth, a remarkable decrease or even no sign of tumor growth was observed (Fig. 7). ...
... Overexpression of ␣ 5  1 integrin into Chinese hamster ovary cells led to loss of tumorigenicity (53). On the other hand, increased expression of ␣ 6 integrin, which involves laminin binding, showed enhanced metastatic potential in Lewis lung carcinoma cells (17). The  1 -subunit associates with at least 10 ␣-subunits (␣ 1 -␣ 9 and ␣ v ), constituting the largest subfamily of the integrins. ...
To investigate the cellular functions of sulfated glycosphingolipids, we introduced the cerebroside sulfotransferase (CST) gene into J5 cells, a subclone of 3LL Lewis lung carcinoma cells. The J5 cells lack acidic glycosphingolipids but accumulate their common biosynthetic precursor, lactosylceramide. We established the stable CST transfectants, J5/CST-1 and J5/CST-2 clones, highly expressing sulfated lactosylceramide (SM3). Both clones exhibited more spherical morphology in comparison to mock transfectant, and their adhesiveness to fibronectin and laminin was significantly lower. The loss of cell-substratum interactions in these SM3-expressing cells could be attributed to decreased expression of integrins (alpha(5), alpha(6), and beta(1)) on the cell surface and their whole cellular levels. However, the levels of H-2K(b) and H-2D(b) antigens remained unchanged. Reverse transcriptase-polymerase chain reaction and Northern blot analyses for these integrins exhibited significant decrease of beta(1) gene expression in J5/CST-1 and 2, but there was no change in the levels of alpha(5) and alpha(6) transcripts. Deglycosylation by endoglycosidase H treatment clearly demonstrated that the precursor form of beta(1) integrin, possessing high mannose oligosaccharide chains, was preferentially decreased in the CST transfectants. These results demonstrate that endogenous SM3 negatively regulates beta(1) integrin expression at the transcriptional level, and the decrease of alpha integrin proteins in the CST transfectants was due to the post-transcriptional modification. We suggest the putative importance of the intracellular pre-beta(1) integrin pool for normal integrin maturation and subsequent function. Although the rates of cell proliferation in vitro for mock and CST transfectants were similar, tumorigenicity of J5/CST-1 and -2 cells inoculated into syngeneic C57/BL6 mice was greatly decreased or even absent. This was probably due to global loss of the efficient cell-matrix interactions, which are essential for the development of malignant tumors in vivo. Thus, we showed the evidence that cellular SM3 negatively regulates the cell-substratum interaction, resulting in the loss of tumorigenicity.
... The expression of certain molecules, such as adhesion receptors and ligands (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) as well as metalloproteinases (18 -22), has been sug-gested to play a role in the development of metastatic lesions. Metastasis occurs via two distinct pathways, and tumor cells spread through blood and LVs 4 . ...
... It has been clearly established that tumor metastasis involves a series of complex processes: detachment of tumor cells from the primary tumor mass, microinvasion into stromal tissues, intravasation into and extravasation from the lymphatic or blood vessels, and growth in secondary sites. Tumor metastasis is now also thought to be associated with dysregulation of cell adhesion, cell motility, and enzymatic proteolysis (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). Various cell adhesion molecules, including E-cadherin, a number of integrins, and CD44, have been suggested as being involved in metastatic processes as a result of experimental studies both in vitro and in vivo as well as descriptive studies using clinical specimens of various types of human cancers in vivo (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)44). ...
... Tumor metastasis is now also thought to be associated with dysregulation of cell adhesion, cell motility, and enzymatic proteolysis (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22). Various cell adhesion molecules, including E-cadherin, a number of integrins, and CD44, have been suggested as being involved in metastatic processes as a result of experimental studies both in vitro and in vivo as well as descriptive studies using clinical specimens of various types of human cancers in vivo (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)44). The findings presented here, however, indicate that differences in the expression of any of these molecules between LNM35 and parental H460 cannot account for the highly metastatic potential of LNM35. ...
Lymphogenous metastasis is a common feature of human lung cancers, but very little is known about the underlying mechanism. In the present study, in vivo selection was carried out to obtain a highly lymphogenous metastatic subline of a human large cell carcinoma of the lung, NCI-H460. The resulting subline, termed NCI-H460-LNM35 (LNM35), was shown to metastasize to regional lymph nodes with a 100% incidence not only as a result of orthotopic intrabronchial (i.b.) implantation, but also as a result of conventional s.c. implantation. LNM35 has a short latency period, allowing for the collection of experimental data within 28 days after i.b. inoculation and 45 days after s.c. inoculation. It was noted that orthotopically i.b.-propagated LNM35 closely mimicked the clinical manifestations of human lung cancer patients by infiltrating into lymphatic vessels and metastasizing to the mediastinal lymph nodes. The LNM35 cell line is, to the best of our knowledge, the first human lung cancer cell line to be reported as having lymphogenous metastatic properties, and the observed 100% incidence by s.c. inoculation gives LNM35 a significant advantage even over previously reported human cancer cell lines of other origins. Comparisons between LNM35 and its parental NCI-H460 cell lines were also made with regard to expression levels and/or activities of various molecules that are thought to play a part in the metastatic process. We show here that the expression of cyclooxygenase 2 is increased in LNM35 and that a specific cyclooxygenase 2 inhibitor, nimesulide, can inhibit the invasion of LNM35 in vitro through Matrigel containing basement membrane components.
Background/Aims: The α6-containing integrin was suggested to be involved in the process of tumor invasion and metastasis. Therefore, the aim of the study was to investigate the expression and function of this adhesion receptor in pancreatic carcinoma. Methods: Integrin expression was investigated in pancreatic tissue and tumor cell lines using immunohistochemistry. Radioimmunoprecipitation was used to determine the complex composition of α6. To analyze the function of the α6-containing integrin in pancreatic cancer, in vitro adhesion, migration, and invasion experiments were performed. Results: The α6-containing integrin was differentially expressed in normal pancreas and pancreatic carcinoma. Immunoprecipitation of different pancreatic carcinoma cell lines showed that α6 was expressed together with the β4 subunit as α6β4 complex. However, adhesion of pancreatic cancer cells to laminin could be inhibited with anti-α6 and anti-β1 integrin antibodies but not by anti-β4 integrin antibody. Migration of the cells through laminin was almost completely inhibited by anti-β1 antibody but not by other anti-integrin antibodies. Tumor cell invasion through a reconstituted basement membrane was only slightly inhibited by anti-α6 antibody. In contrast, a marked inhibition was observed using anti-β1 antibodies, anti-α2-anti-α5 antibodies, and RGDS. Conclusions: The α6-containing integrin is a laminin adhesion receptor in pancreatic carcinoma cells, possibly involved in tumor invasion through the basement membrane.
