野菊花提取物对人肝癌MHCC97H细胞增殖和凋亡的影响

目的 观察中药野菊花提取物(chrysanthemum indicum extact,CIE)对人肝癌MHCC97H 细胞增殖和凋亡的影响及其作用机制.方法 体外培养人肝癌MHCC97H细胞,以0.4、0.8、1.2 mg/ml的CIE(实验1、2、3组)作用于MHCC97H细胞,并设立空白对照组.分别于药物作用24、48、72 h时,应用MTT法检测细胞增殖情况.于药物作用24 h时,用AnnexinV-FITC/PI双染检测细胞凋亡、Rhol23检测细胞线粒体膜电位(△Ψm)变化;Western blot检测细胞色素C、Caspase-9、-3的蛋白表达.结果 三种浓度的CIE对MHCC97H细胞的增殖均有明显的抑制作用,且呈明显的时间和浓度依赖性.与对照组相比,各实验组MHCC97H细胞的凋亡率明显增高(P＜0.05),线粒体膜电位水平明显降低(P＜0.05),细胞色素C、Caspase-9和-3的蛋白表达显著增强(P＜0.05),以上各项均呈明显的量效关系.结论 野菊花提取物对肝癌MHCC97H的增殖具有抑制作用,这可能与药物诱导肝癌细胞凋亡有关.线粒体途径可能是CIE诱导MHCC97H细胞凋亡的主要机制之一.

Effects of Chrysanthemum indicum extract on proliferation and apoptosis of human hepatocellular carcinoma MHCC97H cells

Objective To investigate the effect of Chrysanthemum indicum extract(CIE)on proliferation and apoptosis of hepatocellular carcinoma(HCC)MHCC97H cells and explore its mechanism.Methods MHCC97H cells cultured in vitro,pretreatment MHCC97H cells with CIE 0.4,0.8,1.2 mg/ml and blank control group,proliferation of cells was measured by microculture tetrazolium(MTT).Cell apoptosis and mitochondrial membrane potential(MMP ΔΨm)were determined by flow cytometry.Cytochrome C,Caspase-9 and-3 expression was measured by Western blot.Results CIE significantly inhibited proliferation of MHCC97H cells in a dose-and time-dependent manner.Compared with control group,CIE0.4,0.8,1.2 mg/ml induced MHCC97H cell apoptosis,decreased MMP ΔΨm,and enhanced cytochrome C,Caspase-9 and-3 protein expressions,respectively(P＜0.05).Conclusion CIE can inhibit proliferation of MHCC97H cells,which might be associated with inducing cancer cell apoptosis.Mitochondrial pathway could be one that CIE induce cancer cell apoptosis.