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中药臭灵丹中黄酮类化合物3,5-二羟基-6,7,3′,4′-四甲氧基黄酮(3,5-hydroxy-6,7,3′,4′-tetramethoxyflavone,HTMF)体外对多种肿瘤细胞有抑增殖作用,但机制尚未完全清楚.为探讨HTMF对人鼻咽癌CNE细胞凋亡的影响,用MTT法检测HTMF对CNE细胞株的增殖抑制率,倒置显微镜观察HTMF作用于CNE细胞后细胞形态变化,Hoechst 33258荧光染色观察细胞核形态的变化,流式细胞仪检测细胞的凋亡率,Western blot法检测凋亡蛋白Caspase3和Caspase9的变化,流式细胞仪检测线粒体跨膜电位(△准m)值的改变,并用JC-1荧光染料染色,激光共聚焦显微镜观察线粒体膜电位变化.结果显示,分离自臭灵丹的HTMF呈浓度、时间双重依赖性显著抑制CNE细胞的增殖,诱导细胞凋亡,引起线粒体膜电位下降及凋亡蛋白Caspase3和Caspase9的活化.提示:3,5-二羟基-6,7,3′,4′-四甲氧基黄酮对人鼻咽癌CNE细胞的生长有显著的抑制作用,并可通过降低线粒体膜电位激活Caspase9,进而活化Caspase3诱导CNE细胞凋亡.
Flavonoids in traditional Chinese medicine Silymarin 3,5-dihydroxy-6,7,3 ’, 4’-tetramethoxyflavone (HTMF ) Has inhibitory effect on many kinds of tumor cells in vitro, but the mechanism is not fully understood.In order to investigate the effect of HTMF on the apoptosis of human nasopharyngeal carcinoma CNE cells, MTT assay was used to detect the inhibitory rate of HTMF on CNE cell line by inverted microscopy HTMF cell morphology changes after CNE cells, Hoechst 33258 fluorescence staining observed changes in nuclear morphology, flow cytometry detection of apoptosis rate, Western blot detection of apoptosis protein Caspase3 and Caspase9 changes, flow cytometry mitochondria Transmembrane potential (△ quasi m) value change, and with JC-1 fluorescent dye staining, laser scanning confocal microscope changes in mitochondrial membrane potential.The results show that the concentration of HTMF isolated from stinkyx B in a time-dependent manner significantly inhibited CNE Cell proliferation, induce apoptosis, cause mitochondrial membrane potential decline and activation of the apoptotic proteins Caspase3 and Caspase 9. It is suggested that 3,5-dihydroxy-6,7,3 ’, 4’-tetramethoxyflavone on human Nasopharyngeal carcinoma CNE cells have a significant inhibitory effect, and can be passed Decreasing mitochondrial membrane potential activates Caspase9, which in turn activates Caspase3 to induce CNE cell apoptosis.