蝙蝠葛活性成分对人白血病K562细胞株抗肿瘤作用及机制的研究.docVIP

蝙蝠葛活性成分对人白血病K562细胞株抗肿瘤作用及机制的研究* 杨万山 孙 抒 汪俊颖 全宗学 （延边大学医学院病理学与法医学教研部，吉林 延吉 133000） 摘要：目的：研究蝙蝠葛活性成分体外对人白血病K562S-P法测定凋亡关键效应酶Caspase-9、Caspase-8和Caspase-3的表达情况，深入探讨蝙蝠葛活性成分对人白血病K562细胞株诱导凋亡的机制；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（P0.01）μg/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（P0.01）； 结论：1、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。2﹑蝙蝠葛活性成分对K562细胞抑制增殖作用在20μg/ml作用24h之前主要以诱导凋亡为主。3、蝙蝠葛活性成分诱导凋亡作用机制可能通过上调Caspase-9、spase-8和Caspase-3蛋白表达，经由细胞凋亡的死亡受体通路和线粒体通路完成凋亡的启动和执行。 关键词：蝙蝠葛活性成分；人白血病K562细胞株； Caspase-9蛋白；Caspase-8蛋白；Caspase-3蛋白；细胞凋亡Anti-tumor Action and the mechanism of active components of menispermum dauricum to leukecythemia K562 cells YangWanshan Sun Shu WangJunying QuanZongxue Abstract：Objective: To investigate the effect of active components of menispermum dauricum on inhibiting proliferation and inducing apoptosis of leukecythemia K562 cells.Methods: The growth inhibition rates and cytotoxic effect of leukecythemia K562 cells were measured with MTT 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 leukecythemia K562 cells from active components of menispermum dauricum . Results: (1)MTT assay showed active components of menispermum dauricum could significantly inhibit the growth of leukecythemia K562 cells . The effect of active components of menispermum dauricum was in a dose-dependent manner(P0.01);(2)Under inverted microscope the leukecythemia K562 cells showed the classical apoptosis changes as the cell membrane blurred and apoptotic bodies appeared by treated with 20μg/ml of active 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 leuk