南蛇藤提取物对人胃癌SGC-7901细胞蛋白组学的影响

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。

Effect of Celastrus Orbiculatus Extract on Proteomics of Human Gastric Cancer Cell Line SGC-7901

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 and 48 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 was 70.14 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 and 4 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 COE treatment for gastric cancer.