RNAi对U251细胞c-Met基因表达水平的影响

目的 探讨RAN干扰作用对人脑胶质瘤U251细胞c-Met基因表达的影响。方法 设计3对以c-Met为靶基因短发夹RNA片段，pGenesil-1质粒为载体,构建pGenesil-1/c-Met-shRNA重组质粒，将其转染人脑胶质瘤U251细胞。采用PCR和免疫印迹法分别检测c-Met基因mRNA和蛋白的表达水平。结果 成功构建pGenesil-1-c-Met shRNA重组质粒，并转染至U251细胞。转染重组质粒的U251细胞c-Met基因mRNA和蛋白表达水平均显著降低（P<0.05）。结论 RNA干扰能明显降低U251细胞c-Met基因mRNA和蛋白的表达水平。

Effect of RNA interference on expression of c-Met gene in U251 cells

Objective To construct the shRNA expression vector which targeted c-Met gene and explore the effect of the vector on the expression levels of c-Met gene in the U251 cells. Methods Three pairs of pGenesil-1-c-Met-shRNA recombinant plasmids were constructed. The recombinant plasmids were transfected into the U251 cells. The c-Met mRNA and protein expression levels were determined respectively by the RT-PCR and Western blot methods. Results The pGenesil-1-c-Met shRNA recombinant plasmids were successfully constructed and transfected into U251 cells. The levels of c-Met mRNA expression and protein expression were significantly lower in the U251 cells transfected with pGengsil-1-c Met shRNA recombinant plasmids than those in the U251 cells untransfected with the recombinant plasmids (P<0.05). Conclusion RNA interference can significantly reduce c-Met mRNA and protein expression level in the U251 cells.