HSP90抑制剂17-DMAG对白血病细胞株K562增殖及凋亡的影响

摘要

目的:通过HSP90抑制剂17-二甲基胺乙基-17-去甲氧基格尔德霉素(17-DMAG)作用于白血病K562细胞株,观察HSP90在白血病K562细胞增殖与凋亡中的作用.方法:收集K562细胞,应用HSP90抑制剂17-DMAG作用于K562细胞株,通过半定量PCR检测HSP90的基因表达,WST技术检测17-DMAG对细胞增殖的影响,An-nexin V流式细胞术检测细胞凋亡.结果:17-DMAG处理K562细胞不同时间后,K562细胞的生长明显受抑,且呈时间依赖性(48 h)(r=0.9918)和剂量依赖性(3.2 μmoL/L)(r=0.9999) (P ≤0.01);不同浓度17-DMAG处理K562细胞不同时间后,K562细胞明显凋亡,且呈剂量依赖性(r=0.9903)(P≤0.01);不同浓度17-DMAG作用于K562细胞48 h后,HSP90 mRNA表达明显减少,17-DMAG下调HSP90 mRNA的表达,且呈剂量依赖性(r=0.9227)(P≤0.01).结论:HSP90抑制剂17-DMAG能够抑制白血病K562细胞的增殖,诱导K562细胞凋亡,这为17-DMAG用于白血病的治疗提供实验依据.%Objective:To investigate the role of HSP90 in proliferation and apoptosis of leukemia cells K562 through detecting the effect of HSP90 inhibitors 17-[2-(Dimethylamino) ethyl] amino-17-desmethoxygeldanamycin (17-DMAG) on leukemia K562 cell lines.Methods:The K562 cells were treated with HSP90 inhibitors 17-DMAG,the semi-quantitative PCR was used to detect HSP90 gene expression,the WST was used to detect the effect 17-DMAG on cell proliferation as well as Annexin V flow cytometry was used to detect the cell apoptosis.Results:After 17-DMAG treated the K562 cells in different stage,the K562 cell growth was obviously inhibited with time dependent (48 h)(r =0.9918) and dose dependent(3.2 μmol/L) manners (r =0.9999) (P ≤ 0.01);after the K562 cells in different stage were treated with different concentrations of 17-DMAG,the K562 cells showed significant apoptosis and with dosagedependent mauner (r =0.9903) (P ≤ 0.01);HSP90 mRNA expression decreased significantly after K562 cells were treated with different concentrations of 17-DMAG for 48 hours.17-DAMG down-regulated the HSP90 mRNA expression in dosage-dependent mauner as well(r =0.9227) (P ≤ 0.01).Conclusion:HSP90 inhibitor 17-DMAG can inhibit the proliferation of K562 cells and induce their apoptosis.This study result provides laboratory basis for the treatment of leukemia patients with 17-DMAG.