SmacN7诱导乳腺癌细胞MDA-MB-157凋亡的作用及其机制  

Effects of SmacN7 inducing apoptosis of breast cancer cell MDA-MB-157 and its mechanism

作　　者：吕紫嫣 张肖艳 陈弘 蒋盼若 陈佳玉 LYU Zi-yan;ZHANG Xiao-yan;CHEN Hong;JIANG Pan-ruo;CHEN Jia-yu(Medical school,ShaoxingUniversity,Shaoxing 312000,China)

机构地区：[1]绍兴文理学院医学院,浙江绍兴312000

出　　处：《中国应用生理学杂志》2020年第6期642-647,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(81402979);浙江省科技厅资助项目(2015C33270);浙江省卫生厅资助项目(2014KYA230);国家大学生创新创业训练计划项目(201810349028);浙江省大学生科技创新活动计划项目(2018R432016);绍兴市大学生创新项目(SXSDC201709)。

摘　　要：目的:探讨SmacN7对乳腺癌细胞MDA-MB-157凋亡的作用及机制。方法:将0-20μmol/L的Smac N7应用于乳腺癌细胞MDA-MB-157,用MTS法检测细胞增殖活性,流式细胞仪检测细胞凋亡、细胞周期,hoechst33342染色观察细胞核型变化,JC-1染色检测线粒体膜电位,LDH释放实验检测药物细胞毒性,qPCR检测各基因转录水平,并通过抑瘤实验证实该药抑制乳腺癌增殖的作用。结果:应用Smac N7后,乳腺癌细胞MDA-MB-157增殖抑制率和细胞凋亡率均增加(P<0.01),核型发生显著变化,细胞线粒体膜电位降低,LDH释放量增加,并上调TRAIL、DR4、DR5、p53、PARP-1、Bax、Bid、BAK、caspase-3、caspase-8、caspase-9基因的转录水平(P<0.01),下调Ras、PI3K、AKT、mTOR、Bcl2、Bcl-xL、MCL-1、Survivin、cIAP-1、cIAP-2基因的转录水平(P <0.01)。结论:SmacN7可通过TRAIL介导的死亡受体途径和线粒体介导的内源性凋亡途径诱导乳腺癌细胞MDA-MB-157凋亡,发挥抗乳腺癌的作用。Objective:To investigate the effects and molecular mechanisms of Second mitochondria-derived activator of caspase N7(SmacN7)on the apoptosis of breast cancer cells MDA-MB-157.Methods:Breast cancer cells MDA-MB-157 were treated with SmacN7 at the concentrations of 0-20μmol/L.The proliferation activity of the cells was detected by MTS method,apoptosis and cell cycle were analyzed by flow cytometry,karyotypic changes of MDA-MB-157 cells were observed by Hoechst33342 staining,mitochondrial membrane potential was detected by JC-1 staining,and LDH release experiment was used to detect the drug cytotoxicity.Real time PCR was used to analyze the transcription levels of genes in MDA-MB-157 cells.The effect of inhibiting breast cancer proliferation was confirmed by tumor inhibition experiments.Results:After treated with SmacN7,the inhibition rate of proliferation and apoptosis rate of breast cancer cells MDA-MB-157 were increased(P<0.01),the karyotype changed significantly,the mitochondrial membrane potential in cells was decreased,and the LDH release was increased.The transcription levels of TRAIL,DR4,DR5,p53,PARP-1,Bax,Bid,BAK,caspase-3,caspase-8 and caspase-9 were up-regulated(P<0.01),and the transcription levels of Ras,PI3K,AKT,mTOR,Bcl-2,Bcl-xL,MCL-1,Survivin,cIAP-1 and cIAP-2 were inhibited(P<0.01).Conclusion:SmacN7 induces apoptosis of breast cancer cell MDA-MB-157 through TRAIL-mediated death receptor pathway and mitochondrial-mediated endogenous apoptosis pathway,and plays a role in anti-breast cancer.

关 键 词：SmacN7 乳腺癌 TRAIL通路 线粒体凋亡途径 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]