原花青素B2诱导乳腺癌MCF-7细胞凋亡及其作用机制  

Procyanidin B2-Induced Apoptosis in MCF-7 Cells and Its Mechanism

作　　者：薛宏坤 谭佳琪 李倩[1] 唐劲天[1] XUE Hongkun;TAN Jiaqi;LI Qian;TANG Jintian(Key Laboratory of Particle&Radiation Imaging,Ministry of Education,Department of Engineering Physics,Tsinghua University,Beijing 100084,China;Academy for Advanced Interdisciplinary Studies,Peking University,Beijing 100080,China)

机构地区：[1]清华大学工程物理系,粒子与辐射成像教育部重点实验室,北京100084 [2]北京大学前沿交叉学科研究院,北京100080

出　　处：《食品科学》2021年第9期91-99,共9页Food Science

基　　金：“十二五”国家科技支撑计划项目(2012B1850240)。

摘　　要：本研究旨在探讨原花青素B2(procyanidin B2,PCB2)对人乳腺癌MCF-7细胞的抗肿瘤作用及其机制。采用噻唑蓝法分析PCB2在不同浓度和不同处理时间下对MCF-7细胞存活率的影响;通过激光共聚焦显微镜检测细胞内Ca^(2+)浓度变化并观察Hoechst 33342染色后的细胞凋亡形态;采用流式细胞分析仪检测PCB2对MCF-7细胞凋亡率、周期、胞内活性氧(reactive oxygen species,ROS)水平及线粒体膜电位变化的影响;通过Western blot检测PCB2对MCF-7细胞凋亡相关蛋白表达量的影响。结果表明,当浓度在10~80μmol/L范围内时,PCB2能显著降低MCF-7细胞的存活率(P<0.05),且呈浓度依赖性,处理24、48、72 h时对MCF-7细胞存活率的半抑制浓度分别为114.82、57.15μmol/L和55.64μmol/L。PCB2能显著增加MCF-7细胞凋亡率、胞内ROS水平(P<0.05)、细胞核荧光强度,并破坏Ca^(2+)平衡,同时显著降低线粒体膜电位(P<0.05),并将细胞周期阻滞在G0/G1期进而诱导细胞凋亡。PCB2可上调Bax、细胞色素c、Caspase-12和Caspase-3蛋白的相对表达水平,下调Bcl-2蛋白相对表达水平。综上可知,PCB2具有抗肿瘤作用,其机制与激活线粒体和内质网介导的凋亡通路、周期阻滞和细胞内ROS水平增加有关。The aim of this study was to investigate the anti-tumor effect of procyanidin B2(PCB2)on human breast cancer MCF-7 cells and the underlying mechanism.The methyl thiazolyl tetrazolium method was used to measure the effect of PCB2 at different concentrations and treatment times on the survival rate of MCF-7 cells.The changes in Ca^(2+)concentrations in cells and the apoptotic morphology after Hoechst 33342 staining were observed by confocal laser microscopy.The effects of PCB2 on the apoptosis rate,cell cycle,reactive oxygen species(ROS)level and mitochondrial membrane potential of MCF-7 cells were detected by a flow cytometer.Western blot was employed to determine the effect of PCB2 on the expression levels of apoptosis-related proteins in MCF-7 cells.The results indicated that PCB2 could significantly reduce the survival rate of MCF-7 cells in a dose-dependent manner when the concentration of PCB2 was in the range of 10–80μmol/L.The IC50 for the survival rate of MCF-7 cells at 24,48 and 72 h were 114.82,57.15 and 55.64μmol/L,respectively.PCB2 could remarkably increase the apoptosis rate,ROS level and nuclear fluorescence intensity of MCF-7 cells,destroy the Ca^(2+)balance,dramatically decrease the mitochondrial membrane potential,and block the cell cycle in the G0/G1 phase to induce apoptosis.PCB2 could up-regulate the relative expression levels of Bax,cytochrome c,caspase-12 and caspase-3 proteins,and down-regulate the relative expression level of Bcl-2.In conclusion,PCB2 has antitumor effect,and the underlying mechanism is related to the activation of apoptosis pathway mediated by mitochondria and endoplasmic reticulum,cell cycle arrest and an increase of intracellular ROS levels.

关 键 词：原花青素B2 MCF-7细胞 细胞凋亡 机制 

分 类 号：TS255.42[轻工技术与工程—农产品加工及贮藏工程]