ABSTRACT
OBJECTIVE: To discuss the effect and mechanisms of saikosaponin D (SSD) on apoptosis and autophagy of human hepatocellular carcinoma cells. METHODS: The proliferation of cells in treatment of SSD was detected by MTT. The autophagosome was detected by GFP fluorescence staining. The apoptosis of hepatoma cells were detected by flow cytometry. The expressions of LC3-Ⅱ, p-S6K1 and Beclin 1 in human hepatocellular carcinoma cells was detected by Western blot. RESULTS: The proliferations of PLC, MHCC-97H, SMMC-7721 and Huh7 cells with SSD intervention for 48 h were dose-independent inhibited. The IC50 of PLC, MHCC-97H, SMMC-7721 and Huh7 cells were 51.48, 46.19, 39.87, 48.95 μmol•L-1, respectively. After SSD treatment, the number of autophagosome of PLC, MHCC-97H, SMMC-7721 and Huh7 was significantly more than the normal control group (P<0.05). The apoptosis rate of PLC, MHCC-97H, SMMC-7721 and Huh7 cells induced by SSD+3-MA or SSD+z-DEVD-fmk was less than the SSD treated cells (P<0.05). The expressions of LC3-Ⅱ and phosphorylated S6K1 protein (p-S6K1) in PLC, MHCC-97H, SMMC-7721 and Huh7 cells induced by SSD were more than the normal cells, while the expression of Beclin 1 protein decreased. CONCLUSION: It's suggested that SSD increase the autophagy and apoptosis of human hepatoma cells by regulating the mTORC signaling pathway.