27-O-(E)-香豆酰基-乌索酸通过调控JNK/SAPK通路诱导人乳腺癌细胞MDA-MB-231细胞凋亡  

27-O-( E)-p-coumaric acyl ursolic acid via JNK / SAPK signal pathway regulates apoptosis of human breast cancer MDA-MB-231 cell line

作　　者：王宏婷[1] 王存琴[2] 

机构地区：[1]皖南医学院基础医学院国家中药药理实验室,安徽芜湖241002 [2]皖南医学院药学院,安徽芜湖241002

出　　处：《中国中药杂志》2015年第4期722-726,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81402818);安徽省高等学校省级自然科学研究重点项目(KJ2014A268;KJ2010B246);皖南医学院博士科研启动基金项目(201401)

摘　　要：目的:实验以MDA-MB-231为研究对象,观察27-O-(E)-香豆酰基-乌索酸(DY-17)诱导细胞凋亡作用。方法:采用MTT检测12,24,36,48,60,72 h细胞增殖活性。采用荧光染色技术检测DY-17诱导凋亡实验。运用流式细胞仪测定MDAMB-231的凋亡坏死率。采用Western blotting检测对照组、EGF组、EGF+DY-17 40μmol·L-1组、EGF+SP600125组的JNK,磷酸化JNK,Bax,剪切PARP和剪切caspase-3蛋白表达变化。结果:DY-17抑制MDA-MB-231细胞增殖呈剂量和作用依赖性。流式细胞仪检测DY-17(20,40μmol·L-1)诱导MDA-MB-231细胞凋亡坏死率分别为31.86%,49.91%,荧光显微镜下DY-17(20,40μmol·L-1)与对照组比较细胞发生凋亡坏死明显增加。与对照组比较,EGF组磷酸化JNK蛋白表达上调;与EGF组比较,EGF+DY-17组磷酸化JNK蛋白表达下调。与EGF组比较,EGF+DY-17组Bax,剪切PARP和剪切caspase-3蛋白表达均上调。结论:DY-17通过调控JNK/SAPK通路诱导人乳腺癌细胞MDA-MB-231细胞凋亡。27-O-（E）-p-coumaric acyl ursolic acid（DY-17） from Ilex latifolia is a compound of the monomer. To investigate the DY-17 inducing apoptosis in the human breast cancer cell line, the MDA-MB-231 cells were used as research object in this experiment. The proliferation activity of the MDA-MB-231 cells stimulated with the different concentrations of DY-17 （20, 40 μmolL-1） was detected at different time（12, 24, 36, 48, 60,72 h）. We surveyed the DY-17 inducing apoptosis of the MDA-MB-231 cells with the fluorescent staining technology. The rate of MDA-MB-231 cells apoptosis and necrosis was determined by flow cell cytometry（FCC）. Moreover, expression of JNK, phosphorylated JNK, Bax, PARP shear and caspase-3 shear related to JNK/SAPK pathways were investigated in every group（control group, EGF group, EGF＋DY-17 40 μmolL-1 group and EGF＋SP600125 group）with Western blot. The MTT results showed that, in the presence of DY-17, the proliferation activity of MDA-MB-231 cells decreased in a dose-dependent and time-dependent manner. The apoptosis and necrosis rates of MDA-MB-231 cells with DY-17（20,40 μmolL-1） groups was respectively 31.86%,49.91% by flow cytometry and significantly increased compared with control group under Fluorescence microscopy. Up-regulation of the JNK phosphorylation protein expression was observed in EGF group compared with control group. In addition, markedly decreased the expression of JNK phosphorylation protein were also surveyed in EGF＋DY-17 40 μmolL-1 group compared with EGF group. The expression of Bax, shear PARP and shear caspase-3 protein in EGF＋DY-17 40 μmolL-1 group were significantly increased in comparison with EGF group. The results showed DY-17 induced apoptosis of human breast cancer MDA-MB-231 cell line related to down-regulating JNK/SAPK signal pathways.keywords：27-O-（E）-p-coumaric acyl ursolic acid

关 键 词：27-O-(E)-香豆酰基-乌索酸 MDA-MB-231 JNK/SAPK通路 

分 类 号：R285[医药卫生—中药学]