摘要
目的:探讨南蛇藤提取物(Celastrus orbiculatus extract,COE)在人胃癌细胞中的可能作用靶点。方法:采用细胞活力实验检测COE对细胞生长增殖的抑制作用,计算COE的半数抑制浓度(50%concentration of inhibition,IC50)。蛋白质组学技术寻找COE作用前后的差异表达蛋白;并通过蛋白免疫印迹法(Western blot)对部分蛋白进行验证。结果:COE能够抑制SGC-7901细胞的增殖,呈一定的浓度及时间依赖性;其24 h的IC50为70.14 mg·L^(-1)。COE(70 mg·L^(-1))作用SGC-7901细胞24 h后,与空白组比较有9个蛋白质点发生下调,4个蛋白质点发生上调;这些蛋白可能是COE在胃癌细胞中的作用靶点。Western blot验证了COE对热休克蛋白27(heat shock protein 27,HSP27),抗增殖蛋白(prohibitin),丝切蛋白(cofilin-1),膜联蛋白A5(annexin A5)表达的影响,与蛋白质组学结果相一致。结论:COE对胃癌细胞具有显著的抑制作用,筛选出的差异表达蛋白为COE的抗肿瘤机制研究提供新的依据。
Objective： To investigate the possible effect targets of Celastrus orbiculatus extract（ COE）in gastric cancer cells. Method： SGC-7901 cells were treated with different concentrations of COE for 24 and48 h. The 50% concentration of inhibition（ IC50） of COE was measured by 3-（ 4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide（ MTT） assay. Differentially expressed proteins were detected before and after COE treatment by using proteomic techniques. Partial protein expression was verified by Western blot. Result：COE inhibited the proliferation of SGC-7901 cells in a dose-and time-dependent manner. The IC50 in 24 h was70. 14 mg·L^-1. The protein expression profile showed 13 differentiated expression protein spots after COE（ 70 mg·L^-1） treatment on SGC-7901 cells for 24 h. Among the 13 proteins,9 proteins were down-regulated and4 proteins were up-regulated. These proteins may be the action targets of COE in gastric cancer cells. Its effects on expression of heat shock protein 27（ HSP27）,prohibitin,cofilin-1,and annexin A5 were verified by Western blot,which were consistent with the proteomic results. Conclusion： COE has significant inhibitory effect on gastric cancer cells. The differentially expressed proteins provide new insights into the mechanisms involved in COEtreatment for gastric cancer.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第2期86-90,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81603439)
国家中医药管理局中医肿瘤学重点学科经费项目(国中医药发[2009]30号)
关键词
南蛇藤
胃癌
蛋白质组学
半数抑制浓度
热休克蛋白27
抗增殖蛋白
丝切蛋白
膜联蛋白A5
Celastrus orbiculatus
gastric cancer
proteomic
50% concentration of inhibition（IC50）
heat shock protein 27（HSP27）
prohibitin
cofilin-1
annexin A5
