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目的观察热休克蛋白70基因和蛋白在辛伐他汀诱导K562细胞凋亡过程中的变化,推测热休克蛋白70与细胞凋亡之间的关系。方法20μmol·L-1辛伐他汀处理K562细胞24、48和72h;流式细胞技术检测细胞凋亡率;比色法测定caspase-3蛋白活性;RT-PCR检测caspase-3和热休克蛋白70基因;免疫组织化学法和Western blot技术检测热休克蛋白70蛋白。结果20μmol·L-1辛伐他汀作用K562细胞24、48和72h,细胞凋亡率变化分别是(6.1±s0.4)%,(14.2±0.4)%和(30.7±0.6)%。与相同时间对照组比较,处理组细胞的caspase-3基因和蛋白活性均显著升高(P<0.05);热休克蛋白70的基因和蛋白表达均不同程度下降(P<0.05)。结论辛伐他汀能诱导K562细胞凋亡,降低的热休克蛋白可能是辛伐他汀诱导K562细胞凋亡的机制之一。
Objective To observe the changes of heat shock protein 70 (HSP70) gene and protein during the apoptosis of K562 cells induced by Simvastatin, and to infer the relationship between heat shock protein 70 and apoptosis. Methods K562 cells were treated with 20μmol·L-1 simvastatin for 24, 48 and 72 hours. Flow cytometry was used to detect the apoptosis rate. Caspase-3 protein activity was determined by colorimetric assay. Caspase-3 and heat shock protein 70 Genes; HSP70 protein was detected by immunohistochemistry and Western blot. Results The apoptotic rates of K562 cells treated with 20μmol·L-1 simvastatin for 24, 48 and 72 hours were (6.1 ± s0.4)%, (14.2 ± 0.4)% and (30.7 ± 0.6)%, respectively. Compared with the control group at the same time, the activity of caspase-3 gene and protein in the treated group were significantly increased (P <0.05). The expression of HSP70 gene and protein in the treated group decreased to some extent (P <0.05). Conclusion Simvastatin can induce apoptosis in K562 cells. Reduced heat shock protein may be one of the mechanisms of simvastatin induced apoptosis in K562 cells.