AIF靶向短发夹RNA干扰重组载体对神经母细胞瘤细胞SHSY5Y的沉默作用  

The silent role of AIF short hairpin RNA interference recombinant vector targeting neuroblastoma cells SHSY5Y

作　　者：王占强[1] 李叶丹[2] 李春宇[2] 张极星[3] 王全才[3] 贾春红[1] 张鸿[1] 

机构地区：[1]中国医科大学附属盛京医院神经内科,辽宁沈阳110004 [2]中国医科大学附属盛京医院超声科,辽宁沈阳110004 [3]中国医科大学附属盛京医院神经外科,辽宁沈阳110004

出　　处：《解剖科学进展》2010年第5期409-412,共4页Progress of Anatomical Sciences

基　　金：辽宁省自然科学基金资助项目(No.2008225010)

摘　　要：目的探讨AIF基因短发夹RNA(shRNA)表达质粒对神经母细胞瘤细胞(SHSY5YAIF)基因的干扰作用。方法根据siRNA设计原则,构建靶向AIF基因的短发夹RNA干扰质粒(pGPU6/GFP/AIF),使用脂质体法转染人神经母细胞瘤细胞,通过RT-PCR和Wester blot印迹检测神经母细胞瘤细胞AIF基因mRNA和蛋白表达水平。结果 pGPU6/GFP/AIF重组质粒经限制性内切酶酶切及测序证明基因插入正确。质粒成功转染SHSY5Y细胞后,该细胞的AIFmRNA和蛋白表达明显下降(P<0.05)。结论成功构建靶向AIF基因的shRNA表达载体转染神经母细胞瘤细胞,有效抑制AIFmRNA和蛋白表达,为AIF基因靶向治疗提供前期的实验依据。Objective To investigate gene interference effect of the AIF gene short hairpin RNA(shRNA) expression plasmid on neuroblastoma cells(SHSY5YAIF).Methods According to the siRNA design principles,AIF gene targeting short hairpin RNA interference vector(pGPU6/GFP/AIF) was built,and liposome was transfected into human neuroblastoma cells,AIF gene mRNA and protein expression levels were detected by RT-PCR and Western blot.Results Digesting pGPU6/GFP/AIF recombinant plasmid by restriction enzyme and sequencing proved the correct gene insertion. SHSY5Y plasmid was successfully transfected into SHSY5YAIF cells,AIF mRNA and protein expression in SHSY5YAIF cells was significantly decreased(.P<0.05).Conclusion pGPU6/GFP/AIF targeting AIF gene was successfully constructed and transfected into neuroblastoma cells to inhibit cell AIFmRNA and protein expression,which will provide an initial experimental basis for a target therapy.

关 键 词：神经母细胞瘤 AIF基因 RNA干扰 基因表达 

分 类 号：R739.4[医药卫生—肿瘤]