RNA干扰对MCF-7细胞eIF4E表达的影响

摘要

Aim:To investigate the effect of RNA interfering( RNAi) on the expression of eukaryotic initiation factor 4E(Eif4e) gene in human breast cancer MCF-7 cells. Methods;To design the oligonucleotide contraposed to Eif4e gene, siRNA of Eif4e gene was synthesized by in vitro transcription. Three different concentrations of Eif4e siRNA( 100,150, 200 mg/L) were used to transfect MCF-7 cells in different time periods(24,48,72 h) , and the normal control group, empty liposome group, non-specific siRNA group were established accordingly. The expressions of Eif4e protein and Mrna of each group were detected by using immunocytochemistry and in situ hybridization. Results: After transfecting, in MCF-7 cells,there was a significant decrease in the Eif4e protein and Mrna level in every specific siRNA transfection group of different times, and they were positively correlated with the concentrations of siRNA and the exposure time(P <0.01) ,especially the group of 72 h and the concentration of 200 mg/L(P <0. 05). Conclusion;RNAi can effectively inhibit the expression of Eif4e gene in MCF-7 cells.%目的:探讨RNA干扰(RNAi)对人乳癌MCF-7细胞真核起始因子4E(eIF4E)表达的影响.方法:针对eIF4E基因的寡核苷酸序列设计并体外转录合成小干扰RNA(siRNA),分别以100、150和200 mg/L转染MCF-7细胞24、48及72 h后,采用免疫细胞化学及原位杂交方法检测MCF-7细胞中eIF4E蛋白和mRNA的表达.另设正常对照组(不转染)、空白对照组(转染空脂质体)和无关对照组(转染非特异性siRNA).结果:elF4E siRNA转染MCF-7细胞后,细胞中eIF4E蛋白及mRNA表达量均明显降低,且与转染浓度及时间有一定的依赖关系(P均<0.01),以200 mg/L siRNA转染72 h的抑制效应最为明显(P<0.05).结论:RNAi能有效抑制MCF-7细胞中eIF4E基因的表达.