Rictor-dependent AKT activation and inhibition of urothelial carcinoma by rapamycin

• Published in: Urologic oncology 2012, Vol.30 (1), p.69-77
• Main Authors: Wu, Ming-Ju, M.D., Ph.D, Chang, Chi-Hao, M.Sc, Chiu, Yung-Tsung, Ph.D, Wen, Mei-Chin, M.D, Shu, Kuo-Hsiung, M.D, Li, Jian-Ri, M.D, Chiu, Kun-Yuan, M.D, Chen, Yen-Ta, M.C
• Format: Article
• Language: English
• Subjects: AKT; Animals; Antibiotics, Antineoplastic - pharmacology; Apoptosis - drug effects; Biological and medical sciences; Blotting, Western; Carcinoma, Transitional Cell - drug therapy; Carcinoma, Transitional Cell - metabolism; Carcinoma, Transitional Cell - pathology; Carrier Proteins - metabolism; Cell Proliferation - drug effects; Chromatography, High Pressure Liquid; Disease Models, Animal; Enzyme Activation; Immunohistochemistry; Male; Medical sciences; Nephrology. Urinary tract diseases; Proto-Oncogene Proteins c-akt - metabolism; Rapamycin; Rapamycin-Insensitive Companion of mTOR Protein; Rats; Rats, Inbred F344; Rictor; Sirolimus - pharmacology; Tumors; Urinary Bladder Neoplasms - drug therapy; Urinary Bladder Neoplasms - metabolism; Urinary Bladder Neoplasms - pathology; Urology; Urothelial carcinoma; VEGF
• ISSN: 1078-1439; 1873-2496
• DOI: 10.1016/j.urolonc.2009.11.009

Abstract Objective We previously reported a very high cumulative incidence of urothelial carcinoma in Taiwanese kidney transplant recipients. Rapamycin, the inhibitor of mTOR Complex 1, provides alternative immunosuppressive therapy after kidney transplantation with less neoplastic potential. We examined the in vivo and in vitro effects of rapamycin on urothelial carcinoma. Materials and methods The rat model of urothelial carcinoma was induced by 0.05% N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) in Fischer F344 rats. The anti-tumor effect of rapamycin was assessed grossly, microscopically, and by Western blot analysis. The mechanism of rapamycin's attenuation of urothelial carcinoma was also evaluated by T24 cells. Results Rapamycin significantly reduced urinary bladder tumor growth in the rat model of 0.05% BBN-induced urothelial carcinoma ( P < 0.001). The blood trough levels of rapamycin were correlated with the occurrence of urothelial carcinoma. In vitro, rapamycin also inhibited the cell proliferation, migration, and invasion, as well as the protein expression of vascular endothelial growth factor-A of T24 urothelial carcinoma cells, whereas rapamycin did not induce significant apoptosis in T24 cells. Rapamycin decreased the expression of phospho-mTOR, phospho-S6K, cyclin D1, and VEGF-A. Rapamycin also activated AKT in T24 cells in the rat model of urothelial carcinoma. The rapamycin-associated activation of AKT was inhibited by rictor siRNA, but not raptor siRNA. Conclusions This study provides in vitro and in vivo evidence that rapamycin may inhibit the development of urothelial carcinoma. The present findings also suggest rictor-dependent AKT activation as a consequence of mTORC1 inhibition.