774 DIFFERENTIAL EXPRESSION OF NOTCH-1 AND NOTCH-2 CORRELATES TO INVASIVE PHENOTYPE IN BLADDER CANCER CELL LINES (2010)

You have accessJournal of UrologyBladder Cancer: Basic Research I1 Apr 2010774 DIFFERENTIAL EXPRESSION OF NOTCH-1 AND NOTCH-2 CORRELATES TO INVASIVE PHENOTYPE IN BLADDER CANCER CELL LINES Kilian M. Gust, Shannon Awrey, Susan Ettinger, Estelle Li, Ladan Fazli, and Peter C. Black Kilian M. GustKilian M. Gust More articles by this author , Shannon AwreyShannon Awrey More articles by this author , Susan EttingerSusan Ettinger More articles by this author , Estelle LiEstelle Li More articles by this author , Ladan FazliLadan Fazli More articles by this author , and Peter C. BlackPeter C. Black More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.1423AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The Notch pathway plays a critical role in growth and differentiation, and is involved in cancer progression in different organ sites. Little is known about Notch in bladder cancer. In this study, we evaluated Notch-1 and Notch-2 expression in a panel of bladder cancer cell lines. METHODS Thirteen bladder cancer cell lines with previously described invasive properties and epithelial/mesenchymal characteristics were screened for Notch-1 and Notch-2 expression by Western blot and real-time PCR. Immunohistochemistry (IHC) was performed on a tissue microarray (US Biomax, Inc., Rockville, MD) that included cores from 152 patients with bladder cancer. Alterations in gene expression after Notch-2 silencing with siRNA were measured by expression microarray in 3 cell lines. Genes that changed by at least 2-fold in all 3 cell lines were selected for further analysis. Wound healing assays were performed after treatment with Notch-2 siRNA versus scrambled siRNA. The Mann-Whitney test was performed for statistical analysis. RESULTS Epithelial cell lines with low invasive potential expressed Notch-1 at high and Notch-2 at low relative levels (p = 0.03 and p<0.01, respectively, compared to invasive cell lines). Highly invasive, mesenchymal cell lines expressed high levels of Notch-2 and low levels of Notch-1. These results were validated by Western blot and IHC (TMA). Microarray analysis revealed altered expression of genes related to epithelial to mesenchymal transition (EMT), protein and endosome trafficking, and cell-cell connections, when Notch-2 was silenced with siRNA. Notch-2 silencing decreased the migration of UC3 cells in the wound healing assay (P=0.02). CONCLUSIONS These results provide preliminary evidence for a role for Notch in the progression of bladder cancer and it is suggested that this may be related to EMT. Further investigation is needed to assess the suitability of Notch as a therapeutic target, and it would appear that selective inhibition of individual Notch receptors may be preferential over global inhibition. Vancouver, Canada© 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e302-e303 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kilian M. Gust More articles by this author Shannon Awrey More articles by this author Susan Ettinger More articles by this author Estelle Li More articles by this author Ladan Fazli More articles by this author Peter C. Black More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...