蝙蝠葛活性成分对人白血病K562细胞株抗肿瘤作用的研究

Studies on the Anti- tumor Action of active components of Menispermum dauricum to leukemia K562 cells

目的研究蝙蝠葛活性成分体外对人白血病K562细胞株的抑制增殖和诱导凋亡作用。方法应用MTT法检测蝙蝠葛活性成分对体外培养的人白血病K562细胞株的抑制增殖作用的影响及细胞毒活性;通过倒置显微镜对细胞进行形态学观察;通过免疫细胞化学S-P法测定凋亡关键效应酶Caspase-9、Caspase-8和Caspase-3的表达情况。结果(1)MTT比色法结果表明,蝙蝠葛活性成分对人白血病K562细胞株的增殖抑制作用呈剂量依赖性(P<0.01);(2)倒置显微镜观察:蝙蝠葛活性成分20μg/ml作用后早期细胞出现凋亡形态学变化:细胞膜鼓泡、凋亡小体形成等;作用晚期细胞发生膜破裂坏死;(3)免疫细胞化学结果表明:蝙蝠葛活性成分作用后加药组Caspase-9、Caspase-8和Caspase-3表达增加,与对照组相比较均有显著差异(P<0.01)。结论蝙蝠葛活性成分对人白血病K562细胞株具有抑制增殖和诱导凋亡作用。

Objective To investigate the effect of active components of Menispermum dauricum on inhibiting proliferation and in- ducing apoptosis of leukemia K562 cells. Methods The growth inhibition rates and cytotoxic effect of leukemia K562 cells were measured with MTY assay ; Observing cells morphologic changes under inverted microscope. The Caspase - 9, Caspase - 8 and Caspase - 3 proteins were determined by immunohistochemical S - P staining technique. We explored the mechanism of inducing apoptosis on leukemia K562 cells from active components of Menispermum dauricum. Results （ 1 ） MTI＂ assay showed active com- ponents of Menispermum dauricum could significantly inhibit the growth of leukemia K562 ceils. The effect of active components of Menispermum dauricum was in a dose - dependent manner（ P 〈 0.01 ） ; （2） Under inverted microscope the leukemia K562 cells showed the classical apoptosis changes as the cell membrane blurred and apoptotic bodies appeared by treated with 20/xg/ml of ac- tive components of Menispermum dauricum before 24 hours; after 24 hours cells showed necrosis; （3） Immunocytochemistry showed that the expression of Caspases - 9, Caspases - 8 and Caspases - 3 proteins increased after the leukemia K562 cells treated by active components of Menispermum dauricum, there were different significantly compared with control. Conclusion Active com- ponents of MenisDermum dauricum have the effect of inhibitin~ nroliferation and inducin~ anontosi~ fnr lenkemi~ K562 ~11~.