热休克蛋白90抑制剂格尔德霉素对人肝癌BEL7404细胞株凋亡作用的研究

The effect of geldanamycin, heat shock protein 90 inhibitor, on the apoptosis of human hepatoma cell line BEL7404

目的:研究热休克蛋白90(heat shock protein 90,HSP90)抑制剂格尔德霉素(geldanamycin,GA)对人肝癌细胞株BEL7404的凋亡作用,并探讨其用于肝癌治疗的可行性。方法:体外培养BEL7404细胞株,以GA作用处理;噻唑兰比色(MTT)法检测GA对BEL7404细胞株的增殖抑制作用;吖啶橙荧光染色法及Annexin V-EGFP/PI双染法、透射电镜等方法研究GA对BEL7404细胞株的凋亡作用;流式细胞仪检测BEL7404细胞株的细胞周期变化。结果:GA对BEL7404细胞株具有增殖抑制作用,呈时间剂量依赖关系;GA各剂量组作用24h凋亡率均明显高于阴性对照组(6.31±0.82)%;流式细胞仪检测各剂量组GA均可使BEL7404细胞株细胞周期阻滞于G0/G1期。结论:抑制HSP90的功能可引起肝癌细胞增殖抑制和凋亡,HSP90有可能成为肝癌治疗的新靶点。

Objective To study the effect of geldanamycin（GA）, a kind of heat shock protein 90（HSP 90） inhibitor, on the apoptosis of human hepatoma cell line BEL7404. Methods BEL7404 was cultured and used for GA induction. The cell proliferation, cell cycle and apoptosis of BEL7404 were separately determined by MTT, flow cytometry, acridine orange fluorescent staining and electron microscope assays. Results GA inhibited the proliferation of BELT404 in a dose-and time-dependent manner. The apoptotic level of BELT404 induced by GA was obviously higher than that of the negative control group （P 〈 0.01）. Cell cycle of BEL7404 could be arrestedvat by GA at the G0/G1 phase. Conclusion Inactivation of HSP90 by GA can inhibit cell proliferation and induce apoptosis of hepatoma cells. HSP90 may become a new therapeutic target for hepatocarcinoma.