EGCG对人脑胶质瘤U251细胞SHP1表达的影响

Influence of EGCG on SHP1 expression in human glioblastoma U251 cell line

目的观察表没食子儿茶素没食子酸酯(EGCG)处理后人脑胶质瘤U251细胞SHP1表达及其启动子区甲基化情况的变化,并探讨其对U251细胞增殖的影响及机制。方法以不同浓度的EGCG处理对数生长期的U251细胞,采用CCK-8法检测细胞增殖变化,RT-PCR及Western blotting法检测U251细胞SHP1基因及蛋白表达,MSP法检测EGCG处理后SHP1启动子区甲基化。结果经不同浓度EGCG处理后,U251细胞增殖被抑制,EGCG的抑制作用呈浓度依赖性;U251细胞中SHP1 mRNA的相对表达量及蛋白表达水平明显升高;SHP1启动子区的甲基化水平发生了翻转,由高甲基化转变为低甲基化。结论 EGCG可能通过引起人脑胶质瘤U251细胞SHP1启动子区去甲基化使SHP1表达增加,从而抑制细胞增殖。

Objective To observe the SHP1 expression changes and methylation in the promoter region of human glioblastoma U251 cell line after the treatment of Epigallocatechin Gallate( EGCG),and to investigate the effect and mechanism of EGCG on the cell proliferation of U251 cell line. Methods U251 cells in the logarithmic phase were treated by different concentrations of EGCG. The cell proliferation was detected by CCK-8. The SHP1 gene and protein expression of U251 cells was evaluated by RT-PCR or Western blotting. MSP was used to testify methylation in the promoter region of SHP1 after EGCG treatment. Results After the treatment of different concentrations of EGCG,the proliferation of U251 cell was inhibited,and the inhibitory effect was in a concentration-dependent manner. The SHP1 mRNA and protein expression was elevated in U251 cells,and the promoter methylation of SHP1 was reversed from hypermethylation to hypomethylation. Conclusions EGCG may increase SHP1 expression by causing the methylation in the promoter region of SHP1 of the human glioblastoma U251 cell line,and thus inhibit the proliferation of U251 cell.