Induction Of Apoptosis In Human Liver Carcinoma HepG₂ Cell Line By Casticin

Objective: To investigate the effects of Casticin on the proliferation and apoptosis induction in human hepatocellular carcinoma HepG2 cells, and further to explore the mechanism.Methods : Human hepatocellular carcinoma HepG2 cells were treated with(0,0.5,1.0,2.0)μmol/L Casticin and10μmol/L5-FU, and the cell proliferation inhibition rates of the groups were determined by MTT assay for 12h,24h and 48h after treatment. The Casticin-untrated group is negative controrl group.Morphological changes of cell nucleus were observed by Hoechst 33342 staining. HepG2 cells apoptosis was identified using AV-PI assay by flow cytometry(FCM) and DNA agarosegel electrophoresis. RT-PCR was employ-ed to detect the expression of survivin and Bcl-2 mRNA.Results : It was demonstrated by MTT assay that the growth of HepG2 cells was inhibited by Casticin in a time and concentration-dependent manner (P < 0. 01,P < 0. 05);The rates of inhibition were (38.15±1.72)%,(57.67±0.47)%,(71.88±2.04)% and (53.41±1.34)%,respectively after treatment with various concentration 0.5μmol/L, 1.0μmol/L, 2.0μmol/L Casticin and 10mol/L5-FU for 48h.After treatment with Casticin for 24h,The chromatine concentration and apoptotic cells with high concentrations Casticin were observed by Hoechst 33342 nuclear staining. The percentage of apoptotic cells increa -sed in Casticin-treated groups compared with that in the negative controrl group. Apop -tosis rate of HepG2 cells increased with the increase of concentration detected by flow cytometry(P< 0. 05). A characteristic DNA ladder was demonstrated by DNA agarosegel electrophoresis, indicated DNA break in HepG2 cells.It was indicated by RT-PCR that the expressions of survivin and Bcl-2mRNA in HepG2 cells treated with Casticin was down-regulated.Conclusion:①Casticin siginificantly inhibits the growth of HepG2 cells and induces cell apoptosis in vitro;②The antiproliferation of HepG2 cells may be related to the induction of apoptosis by Casticin ,and down-regalation the expressions of survivin and Bcl-2 mRNA could invole in the effective mechanism of Casticin.