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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。
OBJECTIVE: To investigate the effect of 27-O- (E) -coumaroyl-ursolic acid (DY-17) on the apoptosis of MDA-MB-231 cells. Methods: MTT assay was used to detect the cell proliferative activity at 12, 24, 36, 48, 60 and 72 h. Fluorescent staining was used to detect the apoptosis of DY-17 cells. The apoptosis rate of MDAMB-231 was determined by flow cytometry. The expression of JNK, phosphorylated JNK, Bax, cleaved PARP and cleaved caspase-3 protein in control group, EGF group, EGF + DY-17 40μmol·L-1 group and EGF + SP600125 group were detected by Western blotting. Results: DY-17 inhibited the proliferation of MDA-MB-231 cells in a dose-and dose-dependent manner. The apoptosis necrosis rates of MDA-MB-231 cells induced by DY-17 (20, 40μmol·L-1) by flow cytometry were 31.86% and 49.91%, respectively. DY-17 (20 and 40μmol·L-1) Compared with the control group, apoptosis and necrosis of cells were significantly increased. Compared with the control group, the phosphorylation of JNK protein in EGF group was up-regulated. Compared with EGF group, the phosphorylation of JNK protein in EGF + DY-17 group was down-regulated. Compared with EGF group, the expression of Bax, cleaved PARP and cleaved caspase-3 in EGF + DY-17 group were up-regulated. Conclusion: DY-17 can induce the apoptosis of human breast cancer cell line MDA-MB-231 by regulating the JNK / SAPK pathway.