川楝素诱导人肺癌A549细胞凋亡  

Toosendanin Induces the Apoptosis of Human Lung Cancer A549 Cells

作　　者：刘盛楠[1] 邵淑丽[1] 王维熠 于秋芬[1] 苗长久[1] 李珊珊[1] 冯元[1] 焦凯贺 

机构地区：[1]齐齐哈尔大学生命科学与农林学院,齐齐哈尔161006

出　　处：《中国细胞生物学学报》2015年第8期1087-1094,共8页Chinese Journal of Cell Biology

基　　金：黑龙江省自然科学基金(批准号:C201241);黑龙江省教育厅科学技术项目(批准号:12511611)资助的课题~~

摘　　要：该研究旨在探讨川楝素诱导人肺癌A549细胞凋亡作用及其作用机制。通过不同浓度的川楝素作用于A549细胞48 h后,采用MTT法检测细胞活性;光学显微镜及荧光显微镜下观察细胞形态结构;流式细胞术检测细胞凋亡率、线粒体膜电位(ΔΨm)和细胞周期;实时定量RTPCR和Western blot分别检测Bax、Bcl-2、Fas、Cycs(细胞色素C)和Caspase-3基因m RNA和蛋白质水平。结果显示,在一定浓度范围内,川楝素能抑制A549细胞增殖,诱导细胞凋亡,且呈剂量依赖性。川楝素作用48 h的最佳药物浓度是40μmol/L,增殖抑制率为46.73%±1.47%,细胞凋亡率为13.18%±0.41%,线粒体膜电位(ΔΨm)显著下降(P<0.01),细胞阻滞于G2期和S期;Bcl-2的表达显著降低,Bax、Fas、Cycs和Caspase-3的表达显著增加(P<0.01),提示川楝素可能通过上调Bax、Fas、Cycs和Caspase-3基因和下调Bcl-2基因诱导人肺癌A549细胞凋亡。To investigate the apoptosis-inducing effect of Toosendanin in human lung cancer A549 cells and its mechanisms, MTT assay was used to determine A549 cells activity. Light microscope, fluorescence microscope were used to observe the morphology change of apoptosis in A549 cells. The apoptosis, mitochondrial membrane potential（ΔΨm） and cell cycle distribution of A549 cells were assayed by flow cytometry. The m RNA and protein levels of Bax, Bcl-2, Fas, Cycs（Cytochrome C） and Caspase-3 were measured by Real-time RT-PCR and Western blot. The results showed that Toosendanin inhibited the growth and induced the apoptosis of A549 cells in a dose-dependent manner. The concentration for the treatment was optimized to 40 μmol/L after 48 h in this study and the inhibition rate was up to 46.73%±1.47% for the proliferation, the apoptotic rate could be up to 13.18%±0.41%. The level of ΔΨm decreased and caused a significant S arrest at the expense of G2 phase cell numbers. The results suggested that Toosendanin induced the apoptosis of lung cancer cells through up-regulation of Bax, Fas, Cycs and Caspase-3 and down-regulation of Bc1-2.

关 键 词：川楝素 A549细胞 凋亡 Bcl-2 BAX FAS Cycs CASPASE-3 

分 类 号：R734.2[医药卫生—肿瘤]