摘要
目的探讨α-细辛醚对人食管癌Eca-109细胞线粒体凋亡通路Cyt-c、Smac及Caspase3 mRNA和蛋白表达水平的影响。方法体外培养的人食管癌Eca-109细胞设为阴性对照组和α-细辛醚组,α-细辛醚终浓度分别为25,50,100μg·m L^(-1),培养48 h后使用荧光倒置显微镜观察细胞凋亡形态改变,采用TRIzol法提取细胞总RNA,实时荧光定量聚合酶链反应检测细胞Cyt-c、Smac及Caspase3 mRNA的表达,Western blotting检测其蛋白表达,选取β-actin进行相对定量分析。结果α-细辛醚组细胞形态发生明显凋亡改变;与阴性对照组比较,α-细辛醚组细胞Cyt-c、Smac及Caspase3mRNA和蛋白的表达量均明显增加(P<0.05)。结论α-细辛醚可通过调节线粒体凋亡途径相关基因Cyt-c、Smac及Caspase3的表达增加诱导Eca-109细胞凋亡。
ObjectiveTo discuss the effect ofα-asarone on the expression level of Cyt-c,Smac,Caspase3mRNA and protein in human esophageal carcinoma Eca109cell mitochondria.MethodsThe Eca-109cells were cultured in vitro,and divided into the negative control group and theα-asarone treatment groups(final concentration:25,50,100μg·mL-1).After48h,the morphological changes of Eca109cells were observed by fluorescence inversion microscope.The total RNA of cells were extracted by TRIzol method,the expressions of Cyt-c、Smac and Caspase3were measured by RT PCR and Western blotting.ResultsAfter Eca109cells were treated with different concentrations ofα-asarone for48h,and obvious changes in the morphology were observed,the expressions of Cyt-c,Smac and Caspase3genes and protein were increased significantly compared to the negative control group(P<0.05).Conclusionαasarone can induce the human Eca-109cells apoptosis by regulating expressions of mitochondrial apoptosis pathway correlation genes such as Cyt-c,Smac and Caspase3.
作者
王白燕
张方方
韩倩倩
李瑞琴
朱艳琴
WANG Baiyan;ZHANG Fangfang;HAN Qianqian;LI Ruiqin;ZHU Yanqin(Experimental Teaching Center,Basic Medical School,Zhengzhou 450046,China;Pathology Center,Henan College of Traditional Chinese Medicine,Zhengzhou 450046,China)
出处
《医药导报》
CAS
北大核心
2018年第1期27-30,共4页
Herald of Medicine
基金
郑州市科技攻关项目(131PPTGG417-1)
河南省高等学校重点科研项目(16A310021)
河南省中医药科学研究专项课题(2016ZY2017)
