1134 GRADE ASSOCIATED AUTOPHAGY REGULATION VIA AMPK AND MTOR CONTRIBUTES TO UROTHELIAL CANCER CELL SURVIVAL

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research (III)1 Apr 20131134 GRADE ASSOCIATED AUTOPHAGY REGULATION VIA AMPK AND MTOR CONTRIBUTES TO UROTHELIAL CANCER CELL SURVIVAL Rani U Ojha, Shrawan K Singh, Shalmoli Bhattacharyya, Rakesh Singh, Aruna Rakha, Kusum Joshi, Arup K Mandal, and Vivekanand Jha Rani U OjhaRani U Ojha Chandigarh, India More articles by this author , Shrawan K SinghShrawan K Singh Chandigarh, India More articles by this author , Shalmoli BhattacharyyaShalmoli Bhattacharyya Chandigarh, India More articles by this author , Rakesh SinghRakesh Singh Chandigarh, India More articles by this author , Aruna RakhaAruna Rakha Chandigarh, India More articles by this author , Kusum JoshiKusum Joshi Chandigarh, India More articles by this author , Arup K MandalArup K Mandal Chandigarh, India More articles by this author , and Vivekanand JhaVivekanand Jha Chandigarh, India More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.749AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Urothelial cell carcinoma (UCC) of urinary bladder has high propensity for recurrence and progression. Effective treatment strategies are needed in this direction. Evidences suggest that targeting autophagy may be an useful approach to treat various cancers, however the status and role of autophagy in UCC is largely unknown. To evaluate autophagy in different grades of UCC and autophagy modulators as potential adjunctive therapeutic agent in the management of UCC the study was conducted. METHODS Material consisted of UCC tissue samples from patients with low and high grade tumor, normal urothelial tissues and T24 cell line. The autophagic vesicles (AVs) and expression of various autophagy proteins in tissue samples were studied by transmission electron microscopy and western blotting respectively. Cell viability, mitochondrial membrane potential, ATP, AMPK and mTOR were measured after inducing autophagy by starvation. The therapeutic implication of autophagy was studied using cisplatin alone or in combination with various autophagy inhibitors (AI). RESULTS High grade UCC had more number of AVs and significantly higher expression of autophagic proteins. On starvation primary culture cells derived from high and low grade UCC showed significant induction of autophagy, activation of AMPK and inhibition of mTOR. Increased mTOR activation and decreased autophagic response were observed after AMPK inhibition. Autophagy inhibition by wortmannin, 3-MA and chloroquine led to ATP depletion, mitochondrial membrane hypo-polarization and caspase 3 and 9 dependent cell death. Treatment with cisplatin significantly increased autophagy in UCC. Treatment with combination of cisplatin and AI led to significantly more cell death in UCC as compared to cisplatin or AI alone. All the above findings were related to grade of UCC and were also observed in T24 cells. CONCLUSIONS This novel study shows that autophagy is related to the grade of Urothelial cell carcinoma of bladder and regulated via AMPK - mTOR pathway for tumor cell survival. Autophagy inhibition causes urothelial cancer cell death through intrinsic apoptotic pathway. The potential application of autophagy inhibitors as an adjunct to chemotherapy in UCC needs further exploration. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e463 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rani U Ojha Chandigarh, India More articles by this author Shrawan K Singh Chandigarh, India More articles by this author Shalmoli Bhattacharyya Chandigarh, India More articles by this author Rakesh Singh Chandigarh, India More articles by this author Aruna Rakha Chandigarh, India More articles by this author Kusum Joshi Chandigarh, India More articles by this author Arup K Mandal Chandigarh, India More articles by this author Vivekanand Jha Chandigarh, India More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...