摘要
目的研究青蒿琥酯(Art)对食管癌Ec9706细胞线粒体膜电位的影响,探讨Art诱导食管癌Ec9706细胞凋亡的作用机制。方法以不同浓度(30、60、120μmol/L)的Art作用于食管鳞癌Ec9706细胞12h,对照组以生理盐水代替Art。光学显微镜下观察细胞形态变化,采用流式细胞术Annexin V/PI双染法检测细胞凋亡水平,荧光染料罗丹明123染色检测细胞线粒体膜电位水平。采用流式细胞术检测Caspase-3蛋白表达水平。结果 Art作用后Ec9706细胞形态上呈现不同程度的变化,贴壁细胞变圆,体积变小,飘浮于细胞培养液中。不同浓度Art作用12h后,Ec9706细胞表现出不同程度的细胞凋亡且呈药物剂量依赖性,与对照组相比差异有统计学意义(P<0.01);Ec9706细胞线粒体膜电位表现出不同程度降低且呈药物剂量依赖性,与对照组相比差异有统计学意义(P<0.05)。与对照组相比,Art组Ec9706细胞中Caspase-3蛋白表达水平显著增加(P<0.01),且呈药物剂量依赖性。结论 Art诱导Ec9706细胞凋亡的机制可能是通过降低细胞线粒体膜电位,启动内源性线粒体凋亡途径,激活Caspase-3蛋白,从而诱导细胞凋亡。
Objective To explore the apoptotic mechanism of esophageal cancer Ec9706 ceils induced by artesunate （Art） by means of observing the effect of Art on the membrane potential of Ec9706 cell mitochondria. Methods Ec9706 cells were treated by different doses of Art （30, 60, 120wmol/L） for 12h. The morphological changes in Ec9706 cells were observed by optical microscopy, and the cell apoptosis rate was analyzed by AnnexinV/PI staining. The mitochondrial membrane potential was measured by Rhodamine 123 staining and the expression level of caspase-3 protein was detected by flow cytometry. Normal saline instead of Art was used as control. Results After treatment with Art for 12h, the morphology of Ec9706 cells changed in various degrees, the adherent cells became roundish to a varying degree, and the cells with decreased volume floated in the cell culture medium. After treatment with different doses of Art for 12h, Ec9706 cells showed apoptotic feature to some extent in a dose-dependent manner, and there was statistical significance compared with that of control group （P〈0.01）. The membrane potential of mitochondria in Art group was significantly lower than that in control group （P〈0.05） in a dose-dependent manner. The expression level of caspase-3 protein was significantly higher in Art group than in control group （P〈0.01） in a dose-dependent manner. Conclusion Art may induce the apoptosis of Ec9706 cells by means of reducing the mitochondrial membrane potential, initiating the intrinsic mitochondrial apoptotic pathways and activating caspase-3 protein.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2014年第1期25-29,共5页
Medical Journal of Chinese People's Liberation Army
基金
河北省自然科学基金(H2012206107)
河北省医学科学研究重点课题(20110131)
河北省普通高等学校强势特色学科肿瘤学建设经费[冀教高(2005)52号]~~
关键词
青蒿琥酯
细胞凋亡
膜电位
线粒体
流式细胞术
artesunate
cell apoptosis
potential, mitochondrial membrane
flow cytometry
