RNA干扰抑制STAT6基因表达的初步研究  

The study on gene silence of STAT6 gene by RNA interference

作　　者：刘丹丹[1] 符州[1] 田代印[1] 杨琴[1] 王莉佳[1] 

机构地区：[1]重庆医科大学附属儿童医院呼吸科,重庆400014

出　　处：《重庆医科大学学报》2010年第6期881-884,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:30672268);重庆市卫生局课题(编号:052159)

摘　　要：目的:构建针对信号转导子和转录激活子6(Singnal transducers and activators of transcription 6,STAT6)基因的干扰质粒,并观察其对STAT6蛋白表达的抑制效果。方法:将构建的短发夹干扰质粒(Short hairpin RNA,shRNA)表达载体,与合成的pIRES2-EGFP-STAT6基因表达质粒共同转染COS7细胞,荧光显微镜下观察干扰效果,并利用Western blot观察蛋白表达的变化。结果:将pIRES2-EGFP-STAT6重组载体转染COS7细胞后,结果显示STAT6蛋白可以在细胞中高水平表达;与阴性对照组相比,STAT6 shRNA-1干扰组和STAT6 shRNA-2干扰组STAT6蛋白表达水平分别是23.1%和9.6%。结论:成功构建并筛选到具有显著干扰效率的STAT6干扰质粒,为哮喘的基因治疗研究奠定了基础。Objective：To construct the recombinant plasmid expressing STAT6 shRNA and observe the effects of RNAi suppressed STAT6 gene expression. Methods：shRNA and pIRES2-EGFP-STAT6 plasmid were cotransfected into COS7 cell,to investigate the suppression effect of shRNA on STAT6 under fluorescent microscope . This effect was confirmed by Western blot at protein expression level. Results： STAT6 protein exhibitited high expression in COS7 cells after transfecting pIRES2-EGFP-STAT6 plasmid into COS7 cells alone ; compare with the negative group,the level of the STAT6 protein in the two shRNAs group were 23.1% and 9.6%. Conclusion：STAT6 shRNA were successfully constructed.This technique deserves further study as a potential therapentic tool for asthma .

关 键 词：STAT6基因 STAT6蛋白 RNA干扰(RNAinterference RNAi) SHRNA 哮喘 

分 类 号：R392.12[医药卫生—免疫学]