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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 inhibitory effect of RNAi targeting islet neogenesis related protein (INGAP) on the proliferation of islet cells. Methods siRNA was designed and synthesized and transfected into INS-1 cells by lipofectamine to study its inhibitory effect on target gene. Reverse transcription-polymerase chain reaction (RT-PCR), flow cytometry and Western-blot Method were used to detect the expression of INGAP mRNA and protein in transfected INS-1 cells respectively. The cell proliferation was detected by MTT assay. Results The siRNA6 sequence significantly inhibited the proliferation of cells, the mRNA and protein expression of INGAP was significantly decreased, and the proliferation of INS-1 cells was inhibited (P <0.05). Conclusion Interference of INGAP gene expression can effectively inhibit the proliferation of islet cells. INGAP may be an important new target for the treatment of severe reduction or damage of β cells.