AUTOCRINE IL-6 PRODUCTION BY HUMAN TRANSITIONAL CARCINOMA CELLS UPREGULATES EXPRESSION OF THE α5β1 FIBRONECTIN RECEPTOR

No AccessJournal of UrologyINVESTIGATIVE UROLOGY1 May 2000AUTOCRINE IL-6 PRODUCTION BY HUMAN TRANSITIONAL CARCINOMA CELLS UPREGULATES EXPRESSION OF THE α5β1 FIBRONECTIN RECEPTOR GUANG-JIAN ZHANG, SCOTT A. CRIST, AMY K. MCKERROW, YONG XU, DAVID C. LADEHOFF, and WILLIAM A. SEE GUANG-JIAN ZHANGGUANG-JIAN ZHANG , SCOTT A. CRISTSCOTT A. CRIST , AMY K. MCKERROWAMY K. MCKERROW , YONG XUYONG XU , DAVID C. LADEHOFFDAVID C. LADEHOFF , and WILLIAM A. SEEWILLIAM A. SEE View All Author Informationhttps://doi.org/10.1016/S0022-5347(05)67678-1AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract Purpose: Studies have demonstrated elevated expression and secretion of IL-6 by transitional cell carcinomas (TCC) following bacillus Calmette-Guerin (BCG) therapy. At present, the role of IL-6 on the biology of TCC is poorly understood. This study evaluated the influence of IL-6 expression on a critical variable regulating BCG-tumor interaction, the tumor expression of α5β1 integrin. Materials and Methods: A human TCC cell line (253J) was transfected with an expression vector containing the full-length IL-6 cDNA sequence. Overexpression of IL-6 mRNA and protein was confirmed by Northern analysis and ELISA, respectively. Clones found to overexpress IL-6 were then assayed for α5β1 expression using Northern analysis and flow cytometry. The effect of alterations in α5β1 expression on tumor adherence to fibronectin (FN), and BCG adherence to tumor cells was determined using specific adherence assays. Results: mRNAs for both the α5 and β1 subunits of the FN receptor were increased an average of 9.4 fold and 125.7 fold respectively in the IL-6 overexpressing transfectants relative to the parental 253J cells. Increased mRNA of α5 and β1 was associated with increased cell surface expression of both proteins. Increased protein expression resulted in greater FN substrate binding affinity and increased adherence of BCG to tumor cells. Conclusions: Autocrine expression of IL-6 upregulates the expression of FN receptor subunits in TCC. Increased α5β1 expression increases cellular adherence to FN, and BCG adherence to tumor cells. These results suggest a role for IL-6 in mediating the antitumor activity of BCG by influencing BCG's adherence to TCC. References 1 : Bladder cancer, 1996. CA Cancer J Clin1996; 46: 93. Google Scholar 2 : Intravesical bacillus Calmette-Guerin therapy for murine bladder tumors: initiation of the response by fibronectin-mediated attachment of bacillus Calmette-Guerin. Cancer Res1987; 47: 1762. Google Scholar 3 : Fibronectin mediated Calmette-Guerin bacillus attachment to murine bladder mucosa: requirement for the expression of an antitumor response. J Clin Invest1990; 85: 62. Google Scholar 4 : Role of fibronectin in intravesical BCG therapy for superficial bladder cancer. J Urol1988; 139: 410. Link, Google Scholar 5 : Comparison of the fibronectin-binding ability and antitumor activity of various microbacteria. Cancer Res1990; 50: 3843. Google Scholar 6 : Characterization of soluble fibronectin binding to Bacille Calmette-Guerin. J Gen Microbiol1989; 135: 2735. Google Scholar 7 : Mechanism of interaction of the 85B secreted protein of microbacterium bovis with fibronectin. Infect Immun1993; 48: 28. Google Scholar 8 : Radio-immunoassay detection of interferon-gamma in urine after intravesical Evans BCG therapy. J Urol1990; 144: 1248. Link, Google Scholar 9 : Induction of urinary interleukin-1 (IL-1), IL-2, IL-6, and tumor necrosis factor during intravesical immunotherapy with bacillus Calmette-Guerin in superficial bladder cancer. Cancer Immunol Immunother1992; 34: 306. Google Scholar 10 : Detection of urinary interleukin-2, interleukin-2 receptor, and tumor necrosis factor levels in patients with superficial bladder tumors after intravesical BCG immunotherapy. Urology1994; 43: 187. Google Scholar 11 : Changes in urinary cytokines and soluble intracellular adhesion molecule 1 (ICAM-1) in bladder cancer patients after BCG immunotherapy. Clin Exp Immunol1995; 99: 369. Google Scholar 12 : Evaluation of local immune response after intravesical bacille Calmette-Guerin treatment for superificial bladder cancer. Br J Urol1996; 78: 709. Google Scholar 13 : Role of interleukin-8 in onset of the immune response in intravesical BCG therapy for superficial bladder cancer. Urol Res1997; 25: 31. Google Scholar 14 : Cytokine production by the human bladder carcinoma cell line T24 in the presence of bacillus Calmette-Guerin (BCG). Urol Res1993; 21: 349. Google Scholar 15 : Localization of IL-1, IL-2, IL-4, IL-8 and TNF in superificial bladder tumors treated with intravesical bacillus Calmette-Guerin. J Urol1996; 156: 536. Link, Google Scholar 16 : Interleukin-6 production by bladder tumors is upregulated by BCG immunotherapy. J Urol1995; 154: 572. Link, Google Scholar 17 : Prediction of response to treatment in superificial bladder carcinoma through pattern of interleukin-2 gene expression. J Clin Oncol1996; 14: 1178. Google Scholar 18 : Urinary cytokines during intravesical bacillus Calmette-Guerin therapy for superficial bladder cancer: processing, stability, and prognostic value. J Urol1996; 155: 477. Link, Google Scholar 19 : Response of human NK cells to IL-6 alterations of the cell surface phenotype, adhesion to FN and laminin, and tumor necrosis factor-alpha/beta secretion. J Immunol1993; 150: 4844. Google Scholar 20 : Upregulation of integrin alpha 5 beta 1 expression by interleukin-6 in rabbit corneal epithelial cells. Exp Cell Res1995; 218: 418. Google Scholar 21 : In vitro cultivation of epithelial cells derived from tumors of the human urinary tract. Cancer Res1976; 36: 365. Google Scholar 22 : Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem1987; 162: 156. Google Scholar 23 : In vitro particulate adherence to fibronectin: correlation with in vivo particulate adherence to site of bladder injury. J Urol1992; 147: 1416. Link, Google Scholar 24 : Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods1983; 65: 55. Google Scholar 25 : A novel method for monitoring Mycobacterium bovis BCG trafficking with recombinant BCG expressing green fluorescent protein. Clin Diagn Lab Immunol1996; 3: 761. Google Scholar 26 : Structure and expression of the human B cell stimulatory factor-2 (BSF-2/IL-6) gene. EMBO J1987; 6: 2939. Google Scholar 27 : Bacterial lipopolysaccharide (endotoxin) enhances expression and secretion of beta 2 interferon by human fibroblasts. J Exp Med1987; 166: 1300. Google Scholar 28 : Activation of the human "beta 2-interferon/hepatocyte stimulating-factors/interleukin-6" promoter by cytokines viruses and second messenger agonists. PNAS USA1988; 85: 6701. Google Scholar 29 : Bacterial lipopolysaccharide and inflammatory mediators augment IL-6 secretion by human endothelial cells. J Immunol1989; 142: 144. Google Scholar 30 : Interleukin-6 type cytokines. Ann NY Acad Sci1995; 762: 1. Google Scholar 32 : Interleukin-6 activity in urine and serum in patients with bladder carcinoma. J Urol1992; 148: 791. Link, Google Scholar 33 : BCG-induced interleukin-6 upregulation and BCG internalization in well and poorly differentiated human bladder cancer cell line. Eur Cytokine Network1998; 9: 181. Google Scholar 34 : Antiproliferative effect of IL-6 on transitional cell carcinoma of the bladder. Insight into mechanisms of bacillus Calmette-Guerin immunotherapy. Biochem Soc Trans1997; 25: 270S. Google Scholar 35 : Interleukin-6 functions as an autocrine growth factor in human bladder carcinoma cell lines in vitro. Int J Cancer1997; 72: 149. Google Scholar 36 : Expression of the VLA beta-1 integrin family in bladder cancer. Am J Pathol1994; 144: 1016. Google Scholar From the University of Iowa Department of Urology, Iowa City, Iowa, and the Medical College of Wisconsin Division of Urology, Milwaukee, Wisconsin© 2000 by American Urological Association, Inc.FiguresReferencesRelatedDetailsCited BySecanella-Fandos S, Luquin M and Julián E (2018) Connaught and Russian Strains Showed the Highest Direct Antitumor Effects of Different Bacillus Calmette-Guérin SubstrainsJournal of Urology, VOL. 189, NO. 2, (711-718), Online publication date: 1-Feb-2013.CHEN F, LANGENSTROER P, ZHANG G, IWAMOTO Y and SEE W (2018) Androgen Dependent Regulation of bacillus Calmette-Guerin Induced Interleukin-6 Expression in Human Transitional Carcinoma Cell LinesJournal of Urology, VOL. 170, NO. 5, (2009-2013), Online publication date: 1-Nov-2003.CHEN F, ZHANG G, IWAMOTO Y and SEE W (2018) Bacillus Calmette-Guerin Initiates Intracellular Signaling in a Transitional Carcinoma Cell Line by Cross-Linking α5β1 IntegrinJournal of Urology, VOL. 170, NO. 2, (605-610), Online publication date: 1-Aug-2003.Chen F, Crist S, Zhang G, Iwamoto Y and See W (2018) Interleukin-6 Production by Human Bladder Tumor Cell Lines is Up-Regulated by Bacillus Calmette-Guerin Through Nuclear Factor-κB and Ap-1 Via an Immediate Early PathwayJournal of Urology, VOL. 168, NO. 2, (786-797), Online publication date: 1-Aug-2002.Pook S, Esuvaranathan K and Mahendran R (2018) N-Acetylcysteine Augments The Cellular Redox Changes and Cytotoxic Activity of Internalized Mycobacterium Bovis in Human Bladder Cancer CellsJournal of Urology, VOL. 168, NO. 2, (780-785), Online publication date: 1-Aug-2002. Volume 163Issue 5May 2000Page: 1553-1559 Advertisement Copyright & Permissions© 2000 by American Urological Association, Inc.KeywordsBCGfibronectin receptorbladder neoplasmsinterleukin-6MetricsAuthor Information GUANG-JIAN ZHANG More articles by this author SCOTT A. CRIST More articles by this author AMY K. MCKERROW More articles by this author YONG XU More articles by this author DAVID C. LADEHOFF More articles by this author WILLIAM A. SEE More articles by this author Expand All Advertisement PDF DownloadLoading ...