Wang Hong-ting, Wang Cun-qin
Zhongguo Zhong Yao Za Zhi. 2015 Feb;40(4):722-6.
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 µmol · L(-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 µmol · L(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 µmol · L(-1)) groups was respectively 31.86%, 49.91% by flow cytometry and significantly increased compared with control group under Fluores- cence 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 µmol · L(-1) group compared with EGF group. The expression of Bax, shear PARP and shear caspase-3 protein in EGF + DY-17 40 µmol · L(-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.
来自阔叶冬青的27 - O -（E）-对香豆酰乌索酸（DY - 17）是一种单体化合物。为研究DY - 17诱导人乳腺癌细胞系凋亡的作用，本实验以MDA - MB - 231细胞为研究对象。在不同时间点（12、24、36、48、60、72小时）检测不同浓度（20、40 μmol·L⁻¹）的DY - 17刺激后MDA - MB - 231细胞的增殖活性。采用荧光染色技术检测DY - 17诱导MDA - MB - 231细胞凋亡的情况。通过流式细胞术（FCC）测定MDA - MB - 231细胞的凋亡和坏死率。此外，用蛋白质免疫印迹法检测各组（对照组、表皮生长因子（EGF）组、EGF + 40 μmol·L⁻¹ DY - 17组和EGF + SP600125组）中与JNK/SAPK信号通路相关的JNK、磷酸化JNK、Bax、PARP剪切体和caspase - 3剪切体的表达。MTT结果显示，在有DY - 17存在的情况下，MDA - MB - 231细胞的增殖活性呈剂量和时间依赖性降低。经流式细胞术检测，DY - 17（20、40 μmol·L⁻¹）组MDA - MB - 231细胞的凋亡和坏死率分别为31.86%、49.91%，荧光显微镜下与对照组相比显著升高。与对照组相比，EGF组JNK磷酸化蛋白表达上调。此外，与EGF组相比，EGF + 40 μmol·L⁻¹ DY - 17组JNK磷酸化蛋白表达也明显降低。与EGF组相比，EGF + 40 μmol·L⁻¹ DY - 17组Bax、剪切PARP和剪切caspase - 3蛋白的表达显著增加。结果表明，DY - 17诱导人乳腺癌MDA - MB - 231细胞系凋亡与下调JNK/SAPK信号通路有关。
