Aesculetin On Induction Of The Apoptosis Of Human Liver Cancer BEL-7402 Cells By Mitochondrial Dependent Pathway

ObjectiveTo investigate the effects of aesculetin on the apoptosis of human liver cancer cell BEL-7402 and to explore the molecular mechanism.MethodsThe cell inhibitory rate was observed by MTT assay, cell apoptotic rate and distribution of cell cycle was analyzed by flow cytometry(FCM), cell mitochondrial membrane potential was measured by the method of JC-1 staining.To detect the activities of caspase-3,caspase-8 and caspase-9 by the Microplate Reader. The expression of Bax,Bcl-2,caspase-3,caspase-9were detected by the Western blot.ResultsAesculetin inhibit human liver cancer BEL-7402 cells proliferation in vitro significantly. With the increase of aesculetin( 0.025, 0.05, 0.1, 0.2, 0.4, 0.80.8μg/ml), the inhibition rates were 15.74%, 28.25%, 41.81%, 51.01%,58.13%, 71.51%. The cell apoptotic rate of aesculetin( 0.2, 0.4, 0.8mg/ml)were 11.50%, 20.96%, 58.60%, and arrest cells in S phase. The Mitochondrial membrane potential decreases. The activeity of caspase-3 and caspase-9increases, both showing dose-dependent, While the aactivity of caspase-8dose not shows obvious changes. Bax, caspase-8, caspase- 9 protein expression bootsts and Bcl-2 protein expression drops, both showing does dependent relationship.ConclusionsAesculetin inhibits the proliferation of BEL-7402 in vitro, which probable through the mitochondrial pathway inducing apoptosis.