摘要
目的探讨蝙蝠葛提取物对人肺癌细胞系A549诱导凋亡和抗增殖作用及其机制,为开发抗肿瘤新中药提供实验依据。方法应用MTT法测定蝙蝠葛提取物对人肺癌细胞系A549的生长抑制作用;通过倒置显微镜、光学显微镜观察肿瘤细胞凋亡的形态学变化;采用流式细胞术检测A549细胞的凋亡率;应用免疫组织化学技术检测药物处理前后凋亡相关蛋白酶caspase-3、caspase-8、caspase-9的表达。结果 (1)MTT法检测结果:蝙蝠葛提取物对人肺癌细胞系A549有明显的抑制生长的作用,且呈现出浓度的依赖性;(2)倒置显微镜观察结果:实验组肿瘤细胞体积变小、变圆,核染色质凝集,细胞间连接疏松,贴壁能力减弱;(3)HE染色观察结果:实验组肿瘤细胞体积变小、变圆,核染色质浓缩或染色质块形成,有的细胞膜起泡形成凋亡小体;(4)流式细胞术检测结果显示:蝙蝠葛提取物可以诱导A549细胞发生凋亡,加药组出现亚二倍体峰。结论 (1)蝙蝠葛提取物在体外对人肺癌细胞系A549有显著的诱导凋亡作用;(2)蝙蝠葛提取物诱导凋亡作用机制可能通过上调caspase-3、caspase-8和caspase-9蛋白表达,经由细胞凋亡的死亡受体和线粒体通路完成凋亡的启动和执行;(3)蝙蝠葛提取物具有显著的体外抗肿瘤作用,有望开发成一种新的抗肿瘤药物。
Objective To investigate the effects of menispermum dauricum extract on apoptosis of human lung cancer cell line A549. Methods MTT assay was used to determine the effects of menispermum dauricum extract on the growth inhibition of human lung cancer A549 cells.Morphological changes of the apoptosis of A549 cells were observed by using inverted microscope.Flow cytometry was used to detected apoptotic rate of A549 cells.caspase-3,caspase-8 and caspase-9 proteins were determined by immunocytochemical S-P staining technique. Results MTT test showed that menispermum dauricum extract in concentration of 20,40,80,160 mg/L inhibited A549 cells in a dose and time dependent manner.HE staining showed morphological changes of A549 cells： nuclear chromatin condensation,plasma menbrane bleb formation and the apoptotic body formation.Immunocytochemistry showed that the expression of caspases-3,caspases-8 and caspases-9 proteins in A549 cells increased after treatment of menispermum dauricum extract compared with controls. Conclusions Menispermum dauricum extract can induce apoptosis of human lung cancer A549 cells in vitro,which may be related to up-regulation of caspase-3,caspase-8,caspase-9 expression.
出处
《实用肿瘤杂志》
CAS
北大核心
2011年第4期343-346,共4页
Journal of Practical Oncology
基金
国家自然科学基金资助项目(No.30960121)
吉林省中医药管理局中医药科技项目基金资助(No.08sy5102)
