Figure 2

Inhibition of Akt and mTOR increases the oxaliplatin-induced cytotoxicity in cholangiocarcinoma cell lines. The effects of Akt (LY294002), and mTOR (RAD001) inhibitors on the phosphorylation of Akt and P70S6K in (A) RMCCA1 and (B) KKU100 cells were determined by western blotting. Total Akt and P70S6K were used as loading controls. Cells were treated with 10 μM LY294002, 0.5 μM RAD001 or control vehicle (DMSO) for 24 hours. Representatives from 3 independent experiments are shown. The effects of LY294002 and RAD001 in (C) RMCCA1 and (D) KKU100 cells following treatment with or without oxaliplatin are shown. Cells were treated with 10 μM LY294002, 0.5 μM RAD001 or control vehicle (DMSO), followed by the addition of 0–200 μM oxaliplatin for 48 hours. Cell proliferation was measured by WST-1 and analyzed by spectrophotometric analysis (Absorbance = 450 nm). Results are reported as a percentage inhibition of cell proliferation, where the optical density value from vehicle-treated cells were set as 100% of proliferation and represent the mean ± SE of three independent experiments. (*, p < 0.05 versus the same concentration of oxaliplatin)