title: [Cisplatin enhances apoptosis in bladder cancer cells via autophagy].
abstract: OBJECTIVE
  To monitor the cisplatin-induced autophagy and investigate the function of autophagy in bladder cancer cells.
  
  
  METHODS
  The transmission electron microscope was used to detect autophagic vacuoles and the fluorescence microscope to detect GFP-LC3. The expressions of proteins, such as LC3, PARP, mTOR, P70S6K were analyzed by immunoblotting. Cell viability was analyzed by MTS assay, in which rapamycin was used to inhibit mTOR phosphorylation and enhance autophagy. LC3 expression was knocked down by RNA interference.
  
  
  RESULTS
  In bladder cancer cell T24, autophagic vacuoles were observed by the transmission electron microscope and GFP-LC3 aggregation was viewed by the fluorescence microscope after cisplatin treatment. The LC3-II accumulation was enhanced by cispaltin treatment. Particularly at the concentrations of 50, and 100 μmol/L for 48 h , the gray value of LC3-II/Actin(%) increased 30 and 44, respectively. Cisplatin treatment inhibited the phosphorylation of mTOR/P70S6K, which was most significant at the concentration of 100 μmol/L for 48 h. Cisplatin also induced cell viability loss, which was 12% and 45% at the concentrations of 50, and 100 μmol/L for 24 h. This effect could be enhanced by rapamycin (F=74.890,P<0.01). Furthermore, knocking down LC3 by RNA interference reduced PARP cleavage.
  
  
  CONCLUSIONS
  Cisplatin could induce autophagy in bladder cancer cell T24, which promoted cisplatin-induced apoptosis.
authors.affiliation: Department of Urology, Peking University First Hospital, Beijing 100034, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Yang
authors.forename: Dong-dong
authors.initials: DD
authors.lastname: Yuan
authors.forename: Xue-jun
authors.initials: XJ
authors.lastname: Jiang
authors.forename: Zhi-jun
authors.initials: ZJ
authors.lastname: Xi
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D006801:Humans; D001749:Urinary Bladder Neoplasms / Q000473:pathology*
chemical_list: D002945:Cisplatin