Effect of Yanggan Jiedu Sanjie formula on human hepatocellular carcinoma Bel-7402 cells.

OBJECTIVE: To observe the effect of Yanggan Jiedu Sanjie (YGJDSJ) formula on human hepatocellular carcinoma Bel-7402 cells. METHODS: Bel-7402 cells were treated with YGJDSJ. Cell proliferation was detected by cell counting kit-8 assay. Cell apoptosis was identified by Hoechst 33258 staining and flow cytometric analysis. Cell cycle distribution was quantified by flow cytometric analysis. Caspase activities were measured by commercial kit. Cell senescence was detected by senescence-associated β-galactosidase (SA-β-gal) staining. Protein expression and phosphorylation were identified by Western blot. Protein expression was knocked-down by siRNA. RESULTS: YGJDSJ inhibited proliferation of Bel- 7402 cells in a dose- and time-dependent manner. YGJDSJ induced apoptosis and activated caspase- 3, 8, and 9 in Bel-7402 cells. YGJDSJ-induced apoptosis was completely abrogated by a pan caspase inhibitor, Z-VAD-FMK. YGJDSJ also induced cell senescence, up-regulated cyclin-dependent kinase inhibitor 1a (CDKN1a) and CDKN2a expression and down-regulated retinoblastoma protein (RB) phosphorylation in Bel-7402 cells. Specific knockdown of CDKN1a and CDKN2a significantly reduced YGJDSJ-induce cell senescence in Bel-7402 cells. CONCLUSION: YGJDSJ inhibited cell proliferation, induced caspase-dependent apoptosis and CDKN1a/CDKN2a-RB signalling mediated cell senescence in Bel-7402 cells. Our findings suggest that YGJDSJ might be potential for hepatocellular carcinoma treatment.