新藤黄酸诱导人胃癌MGC-803细胞线粒体途径凋亡的实验研究  

Gambogenic Acid Induces Mitochondria-dependent Apoptosis in Human Gastric Carcinoma Cell Line

作　　者：王训翠[1] 朱国旗[1] 程卉[1] 李庆林[1] 

机构地区：[1]安徽中医药大学省部共建新安医学教育部重点实验室/科研实验中心,安徽合肥230038

出　　处：《中药材》2014年第1期95-99,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(81173600);安徽省自然科学基金(11040606M190)

摘　　要：目的：研究新藤黄酸（Gambogenic acid,GNA）对人胃癌MGC-803细胞凋亡的影响,并探讨其抗肿瘤的作用机制。方法：MTT法检测细胞存活率;Annexin V-FITC/PI染色流式细胞术（FCM）检测MGC-803细胞凋亡率;H2DCFDA探针法流式细胞术检测细胞内活性氧的含量;罗丹明123染色FCM检测细胞线粒体膜电位（△Ψm）的改变;Western blot法检测细胞内P53蛋白的表达变化。结果：新藤黄酸在1.0~12.0μmol/L浓度范围内可抑制人胃癌细胞MGC-803细胞的增殖;增加细胞内活性氧的含量;降低线粒体膜电位;诱导细胞发生凋亡并呈浓度依赖性;Western blotting结果显示,新藤黄酸明显升高MGC-803细胞凋亡诱导基因P53的表达。结论：新藤黄酸可诱导人胃癌MGC-803细胞的凋亡,其机制可能与促进P53介导的线粒体途径依赖的细胞凋亡相关。Objective：To study the effects of Gambogenic acid（ GNA） on the growth of human gastric carcinoma cell line MGC-803 and its underlying mechanisms.Methods：MTT assay was used to measure the cell viability.Apoptosis,mitochondrial membrane potential（ MMP）,reactive oxygen species（ ROS） were detected using flow cytometry method.Among them,Annexin V-FITC/PI double staining was employed in the analysis of apoptosis,Rh123 in analyzing MMP and H2DCFDA in analyzing ROS formation.P53 expression was confirmed by Western blot.Results：4.0 μmol/L GNA inhibited MGC-803 cells growth in a time dependent manner from 24 to 48 h.At the concentration range from 1.0 to 12.0 μmol/L,the inhibitory effect was in a concentration dependent manner.After treatment with 4.0 μmol/L GNA for 48 h,apoptosis was obviously observed as assayed by Annexin V-FITC/PI staining.Importantly,MMP was decreased and ROS formation was increased following GNA treatment.Additionally,P53 expression was up-regulated following 4.0 μmol/L GNA treatment in a time dependent manner.Conclusion：GNA induces mitochondria-dependent apoptosis and increases P53 expression in human gastric carcinoma cell line.

关 键 词：新藤黄酸 MGC-803细胞 线粒体途径 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]