热休克蛋白90抑制NIH3T3成纤维细胞凋亡

HEAT SHOCK PROTEIN90 SUPPRESSES NIH3T3 CELL APOPTOSIS

目的实验应用Hsp90过表达系统观察Hsp90抑制TNFα诱导的细胞凋亡,探讨其可能的作用机制。方法采用电穿孔技术建立稳定过表达Hsp90的细胞克隆,应用激光共聚焦显微镜和流式细胞仪观察TNFα和放线菌酮诱导的细胞凋亡,应用比色分析和Western blotting方法检测caspase的变化。结果1)在稳定过表达Hsp90的NIH3T3细胞中,Hsp90能够抑制TNFα诱导的细胞凋亡。2)在凋亡信号转导通路中,Hsp90作用于caspase-8下游、caspase-3上游。结论1)Hsp90能够在细胞凋亡信号转导通路中发挥负性调节因子的作用。2)Hsp90抑制剂作为抗肿瘤药物的机制之一,可能是通过促进caspase-3活化而促进肿瘤细胞凋亡。

Objective To investigate the suppression of TNFα-induced apoptosis by Hsp90 and examine the underlying mechanism. Methods We used electroporation to establish stable Hsp90 overexpression clones, laser confocal microscopy and flowcytometry to observe apoptosis induced by TNFα and cycloheximide, caspase-8 and caspase-3 colorimetric assay and Western blotting to monitor the change of caspases. Results 1） Hsp90 suppressed TNFα-induced apoptosis in stable Hsp90-overexpressing NIH3T3 cells. 2） Hsp90 functioned at downstream of caspase-8 and upstream of caspase-3 in apoptotic signalling pathways. Conclusion 1） Hsp90 acts as a negative-regulatory factor in apoptotic signalling pathways. 2） As an anti-cancer agent, Hsp90 inhibitor may promote cancer cell apoptosis by activating caspase-3.