17-AAG联合顺铂对人肝癌HepG2细胞增殖和凋亡的影响

Effects of 17-AAG combined with cisplatin on proliferation and apoptosis of human hepatoma cell line HepG2

目的观察热休克蛋白90抑制剂17-AAG联合顺铂(DDP)对人肝癌Hep G2细胞增殖的影响,并探讨其机制。方法将对数生长期的Hep G2细胞分为对照组和A、B、C组。对照组只加培养基。A组加入0.625μmol/L的17-AAG,B组加入1.0μg/m L的DDP,C组加入0.625μmol/L的17-AAG和1.0μg/m L的DDP。培养24、48 h后用MTT法测算A、B、C组细胞增殖抑制率,用流式细胞术测算各组细胞凋亡率,用RT-PCR法检测各组Bax、Bcl-2mRNA。结果 A、B、C组培养48 h时细胞增殖抑制率均高于培养24 h时,P均<0.05;培养时间相同时C组细胞增殖抑制率高于A、B组,P均<0.05。培养48 h后A、B、C组细胞凋亡率和Bax mRNA均高于对照组,Bcl-2 mRNA低于对照组,P均<0.05;C组细胞凋亡率和Bax mRNA均高于A、B组,Bcl-2 mRNA低于A、B组,P均<0.05。结论17-AAG对人肝癌Hep G2细胞具有生长抑制作用,其与顺铂联合有协同作用。其机制可能与上调细胞Bax mRNA、下调Bcl-2 mRNA表达有关。

Objective To explore the effects of 17-AAG combined with cisplatin（ DDP） on the proliferation of human hepatocellular carcinoma Hep G2 cells and its mechanism. Methods Hep G2 cells in the logarithmic phase were divided into the control group and groups A,B and C. The control group was only added with medium. Group A was added with0. 625 μmol / L 17-AAG,group B was added with 1. 0 μg / m L DDP,and group C was added with 0. 625 μg / m L 17-AAG and 1. 0 μg / m L DDP. After being cultured for 24 h and 48 h,the cell proliferation inhibition rates of groups A,B and C were determined by MTT,the apoptosis rate was measured by flow cytometry,and the Bax and Bcl-2 mRNA in each group was measured by RT-PCR. Results The cell proliferation inhibition rates of groups A,B and C which were cultured for 48 h were all higher than those of being cultured for 24 h（ all P〈0. 05）,the cell proliferation inhibition rate of group C was higher than those of groups A and B at the same time of culture（ all P〈0. 05）. After being cultured for 48 h,the apoptosis rate and Bax mRNA in the groups A,B and C were higher than that of the control group,while Bcl-2 mRNA was lower than that of the control group（ all P〈0. 05）; the apoptosis rate and Bax mRNA of group C were higher than those of groups A and B,but the Bcl-2 mRNA was lower than those of groups A and B（ all P〈0. 05）. Conclusions 17-AAG inhibits the growth of human hepatocellular carcinoma Hep G2 cells and has synergistic effect with cisplatin. The mechanism may be related to the up-regulation of Bax mRNA and down-regulation of Bcl-2 mRNA expression.