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目的探讨小分子RNA(siRNA)干扰CASK基因表达对大鼠INS-1细胞胰岛素分泌的影响。方法用脂质体lipofectamine 2000将针对靶基因CASK合成的特异性siRNA片段转染INS-1细胞。实时定量PCR及Western blot检测CASK基因表达变化,进行有效干扰片段的筛选。放射免疫法检测钾离子刺激胰岛素分泌(KSIS)功能的改变。结果转染100nM siRNAs 48h后,INS-1细胞中CASK基因表达明显降低(P<0.01),KSIS功能显著降低(P<0.01)。结论靶向siRNA可以特异性地抑制INS-1细胞中CASK基因表达,从而抑制钾离子诱导的胰岛素分泌。
Objective To investigate the effect of small interfering RNA (siRNA) on the insulin secretion of rat INS-1 cells induced by CASK gene expression. Methods Liposomes were transfected into INS-1 cells with Lipofectamine 2000, a specific siRNA fragment targeting the target gene CASK. Real-time quantitative PCR and Western blot detection of CASK gene expression changes, the effective screening of fragments. Radioimmunoassay was used to detect the changes of potassium ion-stimulated insulin secretion (KSIS) function. Results After transfected with 100nM siRNAs for 48h, the expression of CASK gene in INS-1 cells was significantly decreased (P <0.01) and the function of KSIS was significantly decreased (P <0.01). Conclusion Targeting siRNA can specifically inhibit CASK gene expression in INS-1 cells and thus inhibit potassium-induced insulin secretion.