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

siRNA interference of the INGAP expression inhibited the cell propagation of islet cells lines INS-1 cells

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 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 effects of small interfering RNA （siRNA） on the expression of islet neogenesis associated protein （INGAP） and on cell propagation of INS-1 cells. Methods Various siRNAs against the INGAP gene were designed and transfected into islet cells lines INS-l, respectively . The expression levels of INGAP mRNA and protein were detected by RT-PCR, flow cytometry and Western blotting, respectively. The proliferation of INS-1 cells was detected by MTT. Results Compared with irrelevant siRNA6 control group, empty vector control group and unransfected group, the expression levels of INGAP mRNA and protein were obviously reduced in expermiental groups （all P〈0.05）. The cell proliferation speed significantly increased in transfection group （P〈0.05）. Conclusions siRNA against INGAP could effectively down-regulates INGAP expression in INS-lcell line, and it can inhibit cell proliferation.