Effect of Akt/mTOR inhibition on cisplatin sensitivity and apoptosis in OV433 cells. (A), Increased cisplatin-induced apoptosis by Akt inhibition. OV433-P and -CR cells were treated with cisplatin (50 µM) in the presence or absence of LY294002 (10 µM) for 24 h. Total protein was extracted, and Akt phosphorylation and PARP cleavage were determined by Western blot analysis. (B) Akt inhibition increased cisplatin-induced growth inhibition. Both OV433-P and -CR cells were treated as described in (A) and growth inhibition was determined by MTT assays. (C) Increased cisplatin-induced apoptosis by mTOR inhibition. OV433-P and -CR cells were treated with cisplatin (50 µM) in the presence or absence of rapamycin (50 nM) for 24 h. Total protein was extracted, and the levels of PARP, phosphorylated mTOR and p70S6 K were determined by Western blot analysis. (D) mTOR inhibition increased cisplatin-induced growth inhibition. Both OV433-P and -CR cells were treated as described in (C) and growth inhibition was determined by the MTT assay. Data are representative of three independent experiments. **P < 0.001, statistically significant. Cis, cisplatin; LY, LY294002; Rap, rapamycin.