MP83-15 EFFECT OF GEMCITABINE AND MITOMYCIN ON CANCER STEM CELLS IN UROTHELIAL CARCINOMA CELLS

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology IV1 Apr 2016MP83-15 EFFECT OF GEMCITABINE AND MITOMYCIN ON CANCER STEM CELLS IN UROTHELIAL CARCINOMA CELLS Rani Ojha, Shrawan Singh, Arup K Mandal, and Vivekanand Jha Rani OjhaRani Ojha More articles by this author , Shrawan SinghShrawan Singh More articles by this author , Arup K MandalArup K Mandal More articles by this author , and Vivekanand JhaVivekanand Jha More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2197AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES To investigate the effect of chemotherapeutic agents on the behavior of cancer stem cell (CSC) subsets-side population in primary urothelial carcinoma cells (UCC). METHODS Low-grade urothelial carcinoma (LGUC), non muscle invasive high-grade (HGUC), muscle invasive high grade (MIUC) and bladder cancer cell lines (T24 and UM-UC-3) were used. Normal urothelial cells (NUC) were used as a control. Cytotoxicity of gemcitabine and mitomycin was measured by MTT assay in T24 and UM-UC-3 cells. Stemness properties were measured by isolation and characterization of side population (SP) using hoechst staining, by studying the expression of stemness genes using reverse transcriptase PCR analysis and tumor sphere forming ability. Effect of autophagy inhibition or induction on cell viability was measured by annexin-V/PI staining and caspase activity. RESULTS The number of SP was significantly higher in MIUCC (11.7%, p<0.05) and HGUCC (8.9%, p<0.05) as compared to LGUCC (4.2%) (Fig1A). SP cells showed more CD44 expression and tumor spheroid forming ability (Fig1B). Following treatment with gemcitabine or mitomycin, significantly increased number of SP cells was found in LGUCC (9.78%), HGUCC (13.56%), MIUCC (20.45%) and bladder cancer cell lines at 24 hours. In NUC only 1.7% SP cells were found and there was no increase in SP cells (Fig1C). Treatment with gemcitabine or mitomycin caused increased expression of stemness marker in SP (p<0.001) of HGUCC, LGUCC and MIUCC as compared to SP of NUC cells (Fig1D, E). SP of UCC cells treated with gemcitabine or mitomycin for 24 hours showed lower cell death as compared to NSP cells. Higher autophagic response was observed in SP of UCC in the presence of chemo-therapeutic agents as compared to NUC (Fig1F,G). Pharmacological inhibition or siRNA-mediated inhibition of autophagy led to decreased number of SP cells (Fig1H) and tumor sphere forming ability (Fig1I) with concomitant increase in caspase-mediated cell death (Fig1 J,K) in SP of UCC. CONCLUSIONS The present study is the first to show that urothelial cancer cells exposed to chemotherapeutic agents increases the stemness properties in UCC and could be the underlying cause of resistance and relapse of the tumor. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1086-e1087 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Rani Ojha More articles by this author Shrawan Singh More articles by this author Arup K Mandal More articles by this author Vivekanand Jha More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...