中药臭灵丹中3,5-二羟基-6,7,3′,4′-四甲氧基黄酮对人鼻咽癌CNE细胞凋亡的影响及机制  

3,5-Hydroxy-6,7,3′,4′-tetramethoxyflavone Isolated From Laggera pterodonta Induces CNE Cell Apoptosis

作　　者：曹长姝[1] 沈伟哉[1] 李药兰[2,3] 王辉[4] 高明雅[1] 

机构地区：[1]暨南大学医学院解剖学教研室,广州510632 [2]暨南大学药学院中药及天然产物研究所,广州510632 [3]暨南大学中药药效物质基础及创新药物研究所广东省重点实验室,广州510632 [4]暨南大学医学院微生物及免疫学教研室,广州510632

出　　处：《生物化学与生物物理进展》2011年第3期254-261,共8页Progress In Biochemistry and Biophysics

基　　金：广东省自然科学基金(9151064101000027);广东省自然科学基金团队(039213)资助项目~~

摘　　要：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.3, 5-Hydroxy -6, 7, 3＇, 4＇-tetramethoxyflavone （HTMF） isolated from Laggera pterodonta is known to have an antiproliferative effects in vitro on human cancer. However, the exact mechanisms retain unclear. HTMF was investigated for its antiproliferative effects on human nasopharygeal carcinoma CNE cells. 3-（4, 5-Dimethylthiazol-2-yl）- 2, 5-diphenyl tetrazolium bromide （MTT） assay was used to deserve the inhibitory effect of HTMF. The changes of the cell and nuclear morphological characteristics were observed under the inverted and fluorescence microscope. The cell apoptosis was displayed by Hoechst 33258 staining and flow cytometry （FCM）. The expression of Caspase3 and Caspase9 was detected by Western blotting. The mitochondrial membrane potential were analyzed by FCM and laser confocal microscope with Jc-1 fluorescence staining. MTT assay results show that HTMF significantly inhibited the growth of CNE cells in dose and time dependent manners. The IC5o values of HTMF were 69.02, 28.31 and 3.95 mg/L at 24, 48 and 72 h treatments, respectively. The apoptosis percentage in CNE cells is significantly increased compaired with control group. HTMF with 0, 5.0, 10.0, 20.0 and 40.0 mg/L at 48 h and 40.0 mg/L at 0, 6, 12, 24 and 48 h treatments, respectively increased the expression of Caspase3 and Caspase9 and degraded mitochondriaI membrane potential in dose and time dependent manners. The mechanistic investigation revealed that HTMF has high inhibitory effects on the proliferation of CNE cells and induced the apoptosis of CNE cells by the decrease in mitochondrial membrane potential and increasing the expression of Caspase3 and Caspase9.

关 键 词：3 5-二羟基-6 7 3’ 4’-四甲氧基黄酮(HTMT) CNE细胞 细胞凋亡 线粒体 

分 类 号：R739.6[医药卫生—肿瘤] R285.5[医药卫生—临床医学]