表柔比星+5-氟尿嘧啶、奥沙利铂对人肝癌HepG2细胞增殖及凋亡的影响

Effects of epirubicin combined with 5-fluorouracil and oxaliplatin on proliferation and apoptosis of human hepatoma HepG2 cells

目的观察表柔比星、奥沙利铂、5-氟尿嘧啶(5-FU)三种药物联合对人肝癌细胞株HepG2生长的影响,并探讨其可能发生的机制。方法设立空白对照组、表柔比星组、5-FU+奥沙利铂组、表柔比星+奥沙利铂+5-FU组。经MTT实验方法确认各药物48 h的IC50做为联合用药的浓度分别为表柔比星(3.44μmol/L)、5-FU(1.76 mmol/L)、奥沙利铂(6.89mmol/L),实时荧光定量PCR实验方法测定各组间Bcl-2基因、Bax基因表达的变化,Western blot实验方法测定各组间Bcl-2蛋白、Bax蛋白表达的变化。结果各用药组均够抑制HepG2细胞增殖、诱导细胞凋亡、增加Bax mRNA及Bax蛋白的表达,同时降低Bcl-2mRNA及Bcl-2蛋白的表达,且表柔比星+奥沙利铂+5-FU组的作用优于表柔比星组、奥沙利铂+5-FU组,差异有统计学意义(P<0.01)。结论表柔比星+奥沙利铂+5-FU组能够明显抑制HepG2细胞的增殖,其联合作用可能通过促进Bax基因的表达,抑制Bcl-2基因的表达,从而加速肿瘤细胞的凋亡。

Objective To investigate the influence of oxaliplatin combinated with epirubicine and fluorouracil on proliferation and apoptosis of human hepatocarcinoma cell line HepG2 and its possible mechanism. Methods The HepG2 cells were divided into the blank control group,epirubicin group,5-fluorouracil ＋ oxaliplatin group and epirubicin combined with oxalipain and 5-fluorouracil group. To Confirm the drug IC50 was cnfirmedby MTT assy after withtreament by MTT assay. The concentration is respectively epirubicin（3. 44 mol / L）,5-fluorouracil（1. 76 mmol / L） and oxaliplatin（6. 89 mmol / L）. The changes of Bcl-2 and Bax gene expression was measured by the assay of real-time fluorescent quantitative PCR. The changes of Bax,Bcl-2 protein expression was measured by Western blot method. Results The treatment group could inhibit the proliferation of HepG2 cells and promote its apoptosis. The Bax mRNA and Bax protein expression were increased while Bcl-2 mRNA and Bcl-2 protein expression was decreased in HepG2 cells. The curative effect of epirubicin combined with oxaliplatin and 5-fluorouracil group is better than those of epirubicin and oxaliplatin ＋ fluorouracil group,and the difference was statistically significant（P〈0. 01）. Conclusion Epirubicin combined with oxaliplatin and 5-fluorouracil group could significantly inhibit the proliferation of HepG2 cells.