siRNA干扰CASK表达对INS-1细胞胰岛素分泌的影响

Effect of small interfering RNA targeting CASK on the insulin secretion of pancreatic INS-1 cells

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） targeting CASK on the insulin secretion of pancreatic INS-1 cells. Methods siRNA targeting potassium- stimulated insulin secretion（KSIS） gene was chemically synthesized and then transfected into the rat pancreatic INS-1 cells using lipofectamine 2000. The interfering efficiency of the siRNAs was assessed using RT-PCR and Western blot. The changes of KSIS were detected by radioirnmunoassay. Results After transfected with 100 nmol/L siRNAs for 48 h, the expression of CASK gene in the INS-1 cells was inhibited（P〈0.01） and the KSIS decreasd significantly （ P〈0. 01）. Conclusion The siRNAs targeting CASK mRNA can effectively downregulate CASK expression, which leads to an inhibition of KSIS.