重楼水提物对肝癌细胞HepG2增殖及凋亡的影响

The effect of water extract of Paris Polyphylla on HepG2 cell line proliferation and apoptosis in hepatocellular carcinoma

目的探讨重楼水提物对人肝癌HepG2细胞株增殖和凋亡的影响。方法以体外培养的肝癌HepG2细胞为研究对象,MTT法检测重楼对肝癌HepG2细胞增殖的抑制作用,流式细胞仪Annexin V/PI双染法检测重楼对其凋亡的影响,ELISA法检测Bcl-2、Bax蛋白表达的情况。结果不同浓度重楼水提物能有效抑制HepG2细胞的增殖,且呈时间及浓度依赖性,作用效果在12μg/ml时抑制效果最好(P<0.01)。12μg/ml重楼水提物作用HepG2细胞24h、48h后,结果表明HepG2随着重楼水提物作用时间的延长,细胞凋亡率增加,与阴性对照组相比,差异有统计学意义(P<0.01)。12μg/ml重楼水提物作用HepG2细胞24h后,Bcl-2蛋白表达降低,Bax蛋白表达增加,与阴性对照组相比,差异有统计学意义(P<0.05)。结论重楼水提物能抑制HepG2细胞的体外增殖,抑制作用表现出时效关系,其机制可能与促进细胞凋亡、降低Bcl-2蛋白表达、增加Bax蛋白表达有关。

Objective To investigate the effect of Paris polyphylla on HepG2 cell proliferation and apoptosis in human hepatocellular carcinoma. Methods Hepatocellular carcinoma HepG2 cell lines cultivated in vitro were selected in this study. MTT method was used to detect the water extract of Paris polyphylla inhibition on cell proliferation and apoptotic cells were detected by using flow cytometry Annexin V/PI double stai-ning. The expressions of Bax, Bcl-2 were detected by using HepG2 cells was inhibited by water extract of Paris polyphylla at ELISA method. Results The growth of different concentrations showing a time-and concentration-dependent manner, the maximum inhibition was at concentration of 12μg/ml （ P 〈0.01）. After treated with 12μg/ml water extract of Paris polyphylla for 24 h, 48 h, cell apoptosis was induced, with the prolongation of time, apoptosis increased. Compared with negative control group, cell apoptosis increased sig-nificantly （ P %0.01）. After treated with 12 μg/ml water extract of Paris polyphylla for 24h, the expression of Bcl-2 protein in HepG2 cells decreased, and the expressions of Bax protein increased. Compared with negative control group, there was statistically significant difference （ P 〈0.05）. Conclusion Water extract of Paris polyphylla can inhibit HepG2 cells proliferation in hepatocellular carcinoma with time-effect relationship. The mechanism might be related to inducing cell apoptosis, decreasing expression of Bcl-2, and increasing expression of Bax.