大黄素通过内源性线粒体途径和外源性死亡受体途径诱导人宫颈癌Hela细胞凋亡的研究  

STUDY ON INDUCTION OF EMODIN ON APOPTOSIS OF HUMAN CERVICAL CANCER HELA CELLS VIA INTRINSIC MITOCHONDRIAL AND EXTRINSIC DEATH RECEPTOR PATHWAY

作　　者：王耀先[1,2] 郝慧南 陈秀玮[1] 于辉[1] 娄阁[1] 吴效科[2] 

机构地区：[1]哈尔滨医科大学附属第三医院,哈尔滨150080 [2]黑龙江中医药大学附属第一医院,哈尔滨150040 [3]大庆市第五医院,大庆163714

出　　处：《中国中医药科技》2013年第1期37-39,6,共3页Chinese Journal of Traditional Medical Science and Technology

基　　金：哈尔滨市科技创新人才研究专项资金项目No.2012RFLXSO15

摘　　要：目的:观察大黄素对Hela细胞增殖的抑制作用并探讨其作用机制。方法:倒置显微镜观察细胞形态变化,MTT法检测大黄素对Hela细胞增殖的抑制情况,流式细胞仪检测Hela细胞凋亡,Westernblot检测Hela细胞中凋亡相关蛋白Cytochomec、Apaf-1、Fas、FasL、FADD的表达。结果:空白组Hela细胞呈现典型的多边形和鹅卵形细胞,胞体丰满,边缘清晰,而大黄素处理过的Hela细胞边缘变钝,出现很多皱缩和细小的碎片,表明大黄素对Hela细胞的生长具有毒性作用。MTT法检测结果显示大黄素对Hela细胞的增殖具有明显的抑制作用,并呈剂量依赖性。流式细胞仪定量检测Hela细胞的凋亡结果显示,大黄素在0、20、40、80μM时诱导Hela细胞凋亡率分别为0.8%、8.2%、22.1%、43.7%。Westernblot法检测结果显示,在内源性线粒体途径中,大黄素能够浓度依赖性地增加Cytochromec和Apaf-1蛋白表达;在外源性死亡受体途径中大黄素能够浓度依赖性地增加Fas、FasL和FADD表达。结论:大黄素通过内源性线粒体途径和外源性死亡受体途径诱导Hela细胞凋亡。Objective:To observe the inhibition of emodin on proliferation of human cervical cancer cell HeLa and discuss its mechanism. Methods:The morphology changes of HeLa cells were observed by inverted microscope, the inhibition of emodin on proliferation of HeLa cells was determined by MTT method, apoptosis of HeLa cells was determined by flow cytometer, the apoptosis-related proteins protein such as Cytochome c, apoptotic protease activating factor 1 (Apaf-1), Caspase-9, Fas, FasL, FADD, Caspase-8, and Caspase-3 were detected by Western blot method. Results:HeLa cells in blank control group took the shape of typical polygon and cobble stone, cells were plump, edge of cell was clear, the edge of cell cultured by emodin became dull, body of cell got shrinkage, cell showed small fragments, these showed that emodin had toxic action on HeLa cells. MTT results showed that emodin could obviously inhibit the proliferation of HeLa cells in vitro, the inhibition effect was in a dose-dependent manner. The determination result of flow cytometer showed that emodin could induce apoptosis of HeLa cells, the apoptosis rate of cells respectively was 8.2%,22.1% and 43.7% at concentration of 20,40,80uM.Western blot results showed that emodin could increase Cytochome c and apoptotic protease activating factor 1 (Apaf-1) expression on intrinsic mitochondrial pathway, emodin could enhance Fas, FasL, and FADD expression on dose-dependent manner on extrinsic death receptor pathway. Conclusion:Conclusion : Emodin have toxic action on growth of HeLa cells, dose-dependently inhibit its proliferation and induce its apoptosis, its mechanism was related to cause apoptosis of HeLa cells by intrinsic mitochondrial and extrinsic death receptor pathway.

关 键 词：宫颈肿瘤 中医药疗法 大黄素 治疗应用 细胞凋亡 体外研究 

分 类 号：R285.5[医药卫生—中药学]