白花丹醌对肝癌细胞SMMC-7721凋亡和自噬的影响  

Effect of plumbagin on apoptosis and autophagy of hepatocellular carcinoma cell line SMMC-7721

作　　者：林宇宁 王声善 吕贝贝 马静 陈永欣 钟静 刘雪梅 韦燕飞 LIN Yu-ning;WANG Sheng-shan;LYU Bei-bei;MA Jing;CHEN Yong-xin;ZHONG Jing;LIU Xue-mei;WEI Yan-fei(Department of Physiology,Faculty of Basic Medicine,Guangxi University of Chinese Medicine,Nanning Guangxi 530200,China)

机构地区：[1]广西中医药大学基础医学院生理学教研室,广西南宁530200

出　　处：《中国药理学通报》2018年第10期1419-1424,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金地区项目(No 81560690;81760757);广西自然科学基金资助项目(No 2017GXNSFAA198152);广西中医药大学青年创新研究团队项目(No 2015QT001);广西中医药大学研究生创新课题(No YCSY2018001)

摘　　要：目的研究白花丹醌对肝癌细胞SMMC-7721凋亡和自噬的影响。方法 MTT法检测白花丹醌对SMMC-7721细胞增殖的影响;透射电镜观察细胞凋亡与自噬的形态变化;免疫荧光检测自噬蛋白LC3的变化;qPCR法检测自噬基因Beclin1、Atg7、Atg5 mRNA的变化;Western blot检测自噬蛋白LC3、Beclin1、Atg7、Atg5的表达水平。结果白花丹醌对SMMC-7721细胞增殖有抑制作用,且呈剂量-时间依赖关系;透射电镜下,白花丹醌组细胞凋亡形态明显,并出现自噬体;免疫荧光显示,白花丹醌处理后细胞LC3蛋白的表达上升;qPCR结果表明白花丹醌组自噬基因Beclin1、Atg7、Atg5 mRNA的表达增加;Western blot提示白花丹醌分别与凋亡抑制剂Z-VAD-FMK和自噬抑制剂3-MA联合使用时,能够影响自噬蛋白Atg5、Atg7、LC3-Ⅱ、Beclin1的表达水平。结论白花丹醌促进SMMC-7721肝癌细胞的凋亡与自噬,其机制可能与上调SMMC-7721细胞自噬有关基因及蛋白的表达水平有关。To investigate the effect of plumbagin on the apoptosis and autophagicy of hepatocellular carcinoma cells line SMMC-7721 and its molecular mechanism.Methods The effect of plumbagin on SMMC-7721 cells proliferation was determined by MTT assay.Transmission electron microscopy was used to observe the morphological changes of cell apoptosis and autophagy.Detect tThe changes in autophagy protein LC3 was determined by immunofluorescence.The expression of autophagy genes Beclin1,Atg7 and Atg5 mRNA were measured by qPCR.The expression levels of autophagic proteins LC3,Beclin1,Atg7 and Atg5 were measured by Western blot.Results Plumbagin had certain inhibitory effect on SMMC-7721 cell proliferation,in a dose-and time-dependent fashion.The apoptotic morphology of the plumbagin group was obvious and autophagosomes were present appear.The expression of LC3 protein increased by treatment with plumbagin.The expression of autophagy gene Beclin1,Atg7 and Atg5 mRNA were markedly increased by treatment with concentration of plumbagin.When plumbagin was used in combination with the apoptosis inhibitor Z-VAD-FMK,plumbagin could significantly increase the expression levels of autophagy proteins Atg5,Atg7,LC3-Ⅱand Beclin1.But the expression of autophagic proteins Agt7,Agt5,LC3-Ⅱ,and Beclin1 was significantly decreased in combination with autophagy inhibitor 3-MA.Conclusion The apoptosis and autophagy of SMMC-7721 cells can be promoted by plumbagin,which may be plumbagin raised the level of autophagic gene and protein expression.

关 键 词：白花丹醌 肝癌细胞 凋亡 自噬 LC3 BECLIN-1 Atg7 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]