根皮素诱导肝癌BEL-7402细胞凋亡

目的 研究根皮素诱导肝癌细胞系BEL-7402细胞凋亡.方法 MTT法测定BEL-7402细胞毒性,荧光显微镜观察细胞形态学的变化,流式细胞仪分析细胞周期和线粒体膜电位的变化.发色底物法检测Caspase-3、Caspase-6和Caspase-9活性变化.结果 根皮素对BEL-7402细胞IC50在89.23 μg/mL.BEL-7402细胞生长曲线表明,根皮素浓度增高,生长率明显下降.细胞凋亡可在40～160 μg/mL根皮素处理后24 h出现.凋亡细胞主要表现为核染色质固缩,荧光染色增强.根皮素阻断细胞于G1期,线粒体膜电位降低.Caspase-3、Caspase-6和Caspase-9被激活,呈时间依赖性改变.根皮素处理BEL-7402细胞12 h Caspase-9活性最高,而Caspase-6活性在18 h达峰值,Caspase-3活性峰值时间在24h后.结论 根皮素可以诱导BEL-7402细胞发生凋亡,途径可能是通过线粒体旁路.

Phloretin induces apoptosis of BEL-7402 cells in vitro

OBJECTIVE: To examine the effect of phloretin on apoptosis of BEL-7402 cells. METHODS: The viability changes of BEL- 7402 cells as a result of phloretin-induced toxicity were analyzed using MTT assay, and the cell morphology changes were observed with fluorescence microscope. Flow cytometry was used to analyze the cell cycle and mitochondrial membrane potential changes, and chromogenic substrate assay performed to detect caspase activity. RESULTS: Phloretin induced obvious cytotoxicity against BEL-7402 cells with IC(50) of 89.23 microg/mL. The growth curve demonstrated decreased growth of the cells as phloretin concentration increased. Cell apoptosis occurred 24 h after treatment with 40-160 microg/mL phloretin. Morphological, the cells exposed to phloretin exhibited nuclear chromatin condensation and increased fluorescence intensity. The activity of caspase-9 reached the peak level 12 h after phloretin exposure, and leak levels of caspase-6 and caspase-3 activities occurred 18 and 24 h after the exposure, respectively. CONCLUSION: Phloretin can induce BEL-7402 cell apoptosis though the mitochondrial pathway.