大蓟炭中香叶木素诱导人乳腺癌MCF-7细胞凋亡及其机制研究

摘要

研究大蓟炭中香叶木素诱导人乳腺癌MCF-7细胞凋亡及其作用机制.采用硅胶和Sephadex LH-20柱层析从大蓟炭中分离鉴定了三个黄酮类化合物,经NMR和MS鉴定他们的结构分别为香叶木素(1)、刺槐素(2)和柳川鱼黄素(3).采用MTS方法检测不同浓度的香叶木素对MCF-7的细胞活力的影响;流式细胞术检测不同浓度的香叶木素处理对MCF-7细胞凋亡的作用;Western blot法检测香叶木素处理对细胞PARP、P-JNK等细胞凋亡相关蛋白表达的影响.MTS及流式细胞术结果显示香叶木素能显著抑制MCF-7的增殖并且诱导细胞凋亡;香叶木素可上调P-JNK促进细胞凋亡.结果表明香叶木素在体外实验能通过激活JNK细胞凋亡通路抑制MCF-7的增殖及促进细胞凋亡.%To study the effects of diosmetin from carbonized Cirsium japonicum on apoptosis in human MCF-7 breast cancer cells and its mechanisms.Three flavonoids were isolated and purified from carbonized C.japonicum by silica gel and Sephadex LH-20 chromatography methods.Their structures were elucidated by NMR and MS spectroscopic data and identified as diosmetin(1),acacetin (2) and pectolinarigenin (3).MTS assay was performed to detect the viability of MCF-7 cells treated by different concentrations of diosmetin.The cell apoptosis rate was further analyzed by flow cytometry (FCM).Western blot assay was applied to measure the apoptosis related protein expression levels of PARP,P-JNK.Diosmetin treatment on MCF-7 cells significantly inhibited cell proliferation and induced cell apoptosis.Diosmetin significantly downregulated P-JNK and upregulated cleaved-PARP protein expression.Diosmetin inhibited MCF-7 cell proliferation and induced cell apoptosis by activation of JNK pathway in vitro.