siRNA干扰胰岛新生相关蛋白表达对胰岛细胞株INS-1细胞增殖的抑制作用  

Small interfering RNA-mediated islet neogenesis associated protein gene silencing inhibits the proliferation of INS-1 islet cells

作　　者：沙建平[1] 薛耀明[1] 陈炫[2] 龙可[1] 梁华晟[1] 桑丹[1] 毛睿睿[1] 林占[1] 

机构地区：[1]南方医科大学南方医院内分泌科,广东广州510515 [2]暨南大学第一附属医院临床实验中心,广东广州510630

出　　处：《南方医科大学学报》2009年第10期2040-2043,共4页Journal of Southern Medical University

基　　金："十五"863生物领域高技术首批重点课题(2001AA215161)

摘　　要：目的探讨靶向胰岛新生相关蛋白(INGAP)基因RNAi对胰岛细胞增殖的抑制效应。方法设计合成siRNA,通过脂质体转入胰岛细胞株INS-1中,研究其对靶基因的抑制效应,采用逆转录-聚合酶链反应(RT-PCR)、流式细胞仪、Western-blot法分别检测转染后的INS-1细胞中INGAP mRNA和蛋白表达的变化,用噻唑蓝(MTT)法来检测细胞的增殖。结果结果显示siRNA6序列对细胞增殖有明显的抑制效应,INGAP mRNA及蛋白表达明显降低,INS-1细胞增殖受到抑制,P<0.05。结论干扰INGAP基因的表达能有效抑制胰岛细胞的增殖,INGAP有可能成为治疗β细胞严重减少或损害的重要新靶点。Objective To investigate the effect of small interfering RNA （siRNA）-mediated islet neogenesis associated protein （INGAP） gene silencing on the proliferation of islet cells. Methods Different siRNAs targeting INGAP gene were designed and transfected into INS-1 islet cells, and the expression levels of INGAP mRNA and protein following the transfection were detected using RT-PCR, flow cytometry and Western blotting. The proliferation of the transfected 1NS-1 cells was evaluated using MTT assay. Results Compared with those in the irrelevant siRNA, empty vector control, and un-transfected groups, the expression levels of INGAP mRNA and protein in the cells transfected with siRNA6 were reduced significantly. The cell proliferation rate significantly increased after transfection with siRNA6 （P〈0.05）. Conclusion siRNA targeting INGAP can effectively down-regulate INGAP expression and inhibit the proliferation of INS-1 cells.

关 键 词：RNAI 胰岛新生相关蛋白 细胞增殖 

分 类 号：Q789[生物学—分子生物学]