RNAi对U251细胞c-Met基因表达水平的影响

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

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,p Genesil-1质粒为载体,构建p Genesil-1/c-Met-sh RNA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m RNA和蛋白的表达水平。结果成功构建p Genesil-1-c-Met sh RNA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m RNA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m RNA和蛋白的表达水平。

Objective To construct the sh RNA expression vector which targeted c-Met gene and explore the effect of the vector onthe expression levels of c-Met gene in the U251 cells.MethodsThree pairs of p Genesil-1-c-Met-sh RNA recombinant plasmids wereconstructed. The recombinant plasmids were transfected into the U251 cells. The c-Met m RNA and protein expression levels weredetermined respectively by the RT-PCR and Western blot methods.ResultsThe p Genesil-1-c-Met sh RNA recombinant plasmids weresuccessfully constructed and transfected into U251 cells. The levels of c-Met m RNA expression and protein expression weresignificantly lower in the U251 cells transfected with p Gengsil-1-c Met sh RNA recombinant plasmids than those in the U251 cellsuntransfected with the recombinant plasmids（P0.05）.ConclusionRNA interference can significantly reduce c-Met m RNA and proteinexpression level in the U251 cells.