青蒿素对人白血病细胞Caspase-3及细胞色素C表达的影响

Effect of Artemisinin on Caspase-3 and Cytochrome c of Leuvemia Cell k562

目的:研究青蒿素对人白血病细胞Caspase-3及细胞色素C表达的影响,探讨其对白血病的作用机制,为进一步临床研究及应用提供依据。方法:体外培养K562细胞,用细胞计数法绘制生长曲线;荧光显微镜检测药物作用前后细胞凋亡作用;RT-PCR检测Caspase-3的表达;Western-blot测定药物作用前后线粒体、细胞浆细胞色素C的表达。结果:青蒿素的浓度为1×10-4,1×10-5,1×10-6mol/L时,细胞生长受到显著抑制,并呈剂量依赖性;Hoechst33342/PI双荧光染色可观察到明显的核浓缩、凝集等细胞凋亡表现;RT-PCR检测到Caspase-3的表达;Western-blot检测1×10-5mol/L药物处理细胞后线粒体细胞色素C表达水平下调,细胞浆出现明显细胞色素C蛋白条带。结论:青蒿素能显著抑制人白血病细胞K562的生长,并诱导其凋亡,而促进细胞色素C的释放,激活Caspase-3可能是其作用机制之一。

Objective：To investigate the effect of artemisinin the expression of Caspase-3 and cytochrome c in leukaemia cells k562 also to explore its possible mechanism of anticancer effects.Methods：K562 cells were cultured in vitro.The cell growth curve was drawn according to cell counts.Fluorescence microscopy was applied to demonstrate the presence of apoptosis.The expressins of Caspase-3 were assayed with reverse transcription-polymerase chain reaction（RT-PCR）.The mitochondriaon and cytoplasm of K562 cells treated before and after action by artemisinin were separated and the expression of Cyt c was measured by Western blot.Results：When the concentrations of artemisinin were 1×10^-4,1×10^-5 and 1×10^-6mol/L,the growth of cells was inhibited remarkably in a dose-dependent manner.Fluorescence staining showed obvious apoptosis,such as karyopyknosis and agglutination.RT-PCR assay shows the expression of Caspase-3.Artemisinin could lead to the decrease of mitochondrial cytochrome c concentration,which in turn lead to the increase of cytoplasmic cytochrome c concentration.Conclusion：Artemisinin could inhibit proliferation and induce apoptosis in K562 induction of delivery of cyt-c and activation of Caspase-3 may contribute to its effect.