Figure 2

Concentration-dependent effect of JS-K on ROS, GSH/GSSG ratio, mitochondrial membrane potential, ATP production and apoptotic proteins.

Bladder cancer cells were treated with JS-K (0 μM, 1 μM, 2 μM and 5 μM) for 6 h and then the intracellular level of total ROS (A), the level of superoxide (B), the GSH/GSSG ratio (C), the mitochondrial membrane potential (D), ATP production (E) and caspase-3/7 activity (F) were detected. The ROS levels and caspase-3/7 activity in JS-K-treated cells were significantly increased compared with their levels in the vehicle control. The GSH/GSSG ratio, mitochondrial membrane potential and ATP production were markedly decreased. (G) Cells were treated with JS-K (0 μM, 1 μM, 2 μM and 5 μM) for 24 h and the levels of Bak, Bax, Bcl-2, caspase-9, PARP and HO-1 proteins were examined by western blot. The data indicate that JS-K regulated apoptotic proteins in bladder cancer cells in a concentration-dependent manner. (H) Cytochrome c was detected in the mitochondria and cytosol and AIF was detected in the nucleus. The release of cytochrome c from the mitochondria to the cytosol was demonstrated. The AIF level was upregulated in JS-K-treated cells. The data are presented as the mean ± SD for at least three independent experiments. Single asterisks (*) indicate a significant difference (P < 0.05) and double asterisks (**) indicate an extremely significant difference (P < 0.01).