Effect Of Ginsenoside Compound K On Apoptosis Of T24Human Bladder Cancer Cell Lines

It has been reported that ginsenoside can inhibit cancer cells growth and induce apoptosis. Bladder cancer is one of the most common malignancies and its incidence has recently increased. The mechanism of inhibition of bladder cancer with ginsenoside Compound K (CK) was not clear. The aim of the present study was to investigate effects of ginsenoside CK on apoptosis of T24human bladder cancer cells and to assess apoptotic mechanism in T24cells.The cell viability of T24cell lines with different concentrations and periods were investigated by MTT assay. The results showed ginsenoside CK inhibited T24cell growth in dose-and time-dependent manners. Hoechst33342staining analyzed morphological changes by fluorescence microscope, after treatment of ginsenoside CK for24h, the nucleus was significantly smaller and broken into pieces with granular fluorescence. The ratio of apoptosis were determined by flow cytometry, after incubation for24h, the apoptotic rate of T24cells with20μmol/L CK was more than90%. The expression of apoptosis associated protein Bcl-2, Bax, Caspase-3, Caspase-9, p38MAPK and P-p38MAPK were determined by western blot. The results showed CK treatment resulted in an appreciable up-regulation of protein expression of Bax and P-p38MAPK, down-regulation in Bcl-2, up-regulation of active type of Caspase-3and Caspase-9proteins in T24cells and these effects could be completely blocked by the p38MAPK special inhibitor SB203580.Accumulation of ROS (reactive oxygen species) in T24cells was determined, the results found that the levels of ROS were significantly increased after CK treatment and this effect could be blocked by NAC (N-acetyl-L-cysteine).In conclusion, this study suggested the apoptosis-promoting effects of CK on bladder cancer T24cells were dependent on ROS generation, p38MAPK activation and were associated with the apoptosis proteins of Bcl-2family and caspase family.