热休克蛋白90抑制剂17-AAG对K562/ADR耐药细胞株生长和凋亡的影响

目的研究热休克蛋白90抑制剂17-AAG对耐药白血病细胞生长的影响及其诱导的细胞凋亡作用,为白血病的治疗提供新的研究思路。方法采用MTT法检测17-AAG对细胞增殖抑制的剂量-时间-效应关系;激光共聚焦DA-PI染色法观察细胞形态学变化;流式细胞术检测细胞凋亡、周期及P-糖蛋白(P-gp)的变化,Western blot检测细胞凋亡相关蛋白的变化。结果 17-AAG能够明显抑制K562/ADR细胞的生长,大量细胞的核内DNA发生断裂,细胞核边缘化,加药组细胞凋亡率与对照组相比明显增高;加药后细胞被阻滞在G1期,procaspase-3和cleaved Caspase-3表达升高。结论 17-AAG可以通过诱导耐药细胞株K562/ADR的凋亡而抑制细胞生长。P-gp和Caspase-3的表达与凋亡事件的发生具有一定的相关性。

Effects of heat shock protein 90 inhibitor 17-AAG on the growth and apoptosis in K562/ADR cells

Aim To explore the effects of 17-AAG on the growth and apoptosis of drug-resistant leukemia K562/ADR cells and provide new way for leukemia therapy.Methods K562/ADR cells were treated with 17-AAG.Cell viability was evaluated by MTT assay. 17-AAG induced apoptosis of K562/ADR cells was delineated by DAPI staining assay;the apoptosis,the cell cycle distribution and the expression of P-gp were determined using flow cytometry.Furthermore,the expression diversity of related protein was examined by Western blot assay.Results 17-AAG effectively inhibited the growth of K562/ADR cells;cell cycle was arrested at the stage of G0/G1 and cellular DNA collapse occurred after treating with 17-AAG.Moreover,the expression of procaspase-3 and cleaved Caspase-3 increased in the presence of 17-AAG.Conclusions 17AAG can inhibit the growth of K562/ADR cells and induce apoptosis.P-gp and Caspase-3 may be involved in apoptosis induced by 17-AAG.