HSP90抑制剂GA对人肝癌BEL7404细胞株凋亡作用的研究

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，并探讨相关机制及意义。方法：体外培养BEL7404细胞株，以HSP90抑制剂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：HSP90抑制剂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（5 umol/L、10 umol/L、15 umol/L）作用24h凋亡率（%）分别为10.2±1.35、21.3±1.30、38.7±1.43，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：通过抑制HSP90的功能，GA能明显抑制肝癌细胞增殖，促进肝癌细胞凋亡，使肝癌细胞周期阻滞于G0/G1期，其抗癌活性可能与其抑制HSP90的分子伴侣功能有关。

Influence of heat shock protein 90 inhibitors on apoptosis of human hepatoma cell line BEL7404

AIM: To study influence of geldanamycin（GA）on human hepatoma cell line BEL7404. METHODS: The effect of GA on cell proliferation, cell cycle and apoptosis of BEL7404 were investigated by the methods of MTT, flow cytometry ,acridine orange fluorescent staining and electron microscope. RESULTS:After treatment with GA ,the proliferation of BEL 7404 cells was obviously inhibited in a dose and time –dependent manner. The apoptotic rate (%) of BEL7404 cells which were treated 24h with GA were 10.2±1.35、21.3±1.30、38.7±1.43 , respectively with higher significance than that of the control group (P<0.01).GA could arrest cell cycle to G0/G1 phase. CONCLUSION: GA can induce the apoptosis and arrest cell cycle to G0/G1 phase of BEL7404, and its action mechanism may be correlated with its inhibition of heat shock protein 90 molecular chaperone activity.