Salvianolate induces apoptosis of human hepatoma SMMC-7721 cells through mitochondrial pathway / 中国肿瘤生物治疗杂志

ABSTRACT

Objective: To explore the apoptosis-inducing effect of salvianolate on hepatoma SMMC-7721 cells and the underlying mechanism. Methods: SMMC-7721 cells were co-cultured in vitro with different concentrations (0.5, 1, 2 mg/ml) of salvianolate for 24 h. The apoptotic SMMC-7721 cells were examined by flow cytometry, and the changes of mitochondrial transmembrane potential were examined by mitochondrial transmembrane potential JC-1 kit. The activities of caspase-8, caspase-9, and caspase-3 were detected by spectrophotometry in the hepatoma SMMC-7721 cells after co-cultured with 1 mg/ml salvianolate. The changes of apoptotic SMMC-7721 cells induced by salvianolate in the presence or absence of caspase-9 inhibitor or caspase-3 inhibitor were measured by flow cytometry. The expressions of pro-apoptotic protein Bax and anti-apoptotic protein Bcl-2 were detected by Western blotting analysis. Results: Salvianolate significantly induced apoptosis of hepatoma SMMC-7721 cells (P<0.05), and the decline of mitochondrial membrane potential increased with the increase of salvianolate concentration (P<0.05). The activities of caspase-9 and caspase-3, but not caspase-8, were increased in hepatoma cells after treatment with 1 mg/ml salvianolate for 24 h (P<0.05). The apoptosis-inducing effect of salvianolate was significantly decreased in the presence of caspase-9 or caspase-3 inhibitors (P<0.05). Western blotting results showed that salvianolate increased pro-apoptotic protein Bax expression and decreased anti-apoptotic protein Bcl-2 expression. Conclusion: Salvianolate can induce the apoptosis of human hepatoma SMMC-7721 cells in a dose-dependent manner, which is probably mediated by mitochondrial apoptosis pathway.