PDX-1基因对人胚肾细胞胰岛发育相关基因表达的影响

Effect of PDX-1 on the expression of pancreas islet development-related genes

【目的】研究人PDX-1基因对胰岛发育相关基因和胰岛素基因(Insulin,INS)表达的影响,探讨PDX-1基因诱导非β细胞为胰岛素分泌细胞的可能性。【方法】采用PCR扩增技术,从人的胰腺cDNA中扩增PDX-1基因和INS基因的阅读框架,分别构建了质粒pAAV-PDX-1和pAAV-INS;将pAAV-PDX-1转染人胚肾细胞(293T)48 h后,RT-PCR检测293T细胞中胰岛发育相关基因的表达;再将pAAV-PDX-1和pAAV-INS共转染293T细胞48 h后,RT-PCR检测293T细胞中INS的表达。【结果】转染pAAV-PDX-1的293T细胞中,检测到胰岛发育相关基因PDX-1、Ngn3、PC2、BETA2的表达;PDX-1没有直接增强INS基因表达的作用。【结论】在293T细胞中,PDX-1基因对胰岛发育相关基因的表达有重要影响,PDX-1在诱导非β细胞为胰岛素分泌细胞的过程中可能有着重要的作用。

【Objective】 The research was conducted to study the function of the PDX-1 gene and the effect of PDX-1 gene on regulation of INS gene expression in non-β-cells.【Method】 The plasmid pAAV-PDX-1 was transfected into the 293T cells and the expression of the pancreas islet development-related genes was analyzed by RT-PCR.The plasmid pAAV-PDX-1 and pAAV-INS were transfected into the 293T cells and the expression of the INS gene was analyzed by RT-PCR.【Result】 Some insulin-secreting cell related genes（PDX-1,Ngn3,PC2,BETA2） were expressed in the 293T cells transfected with pAAV-PDX-1.There was no detectable change in the expression of the INS gene in the 293T cells co-transfected with pAAV-PDX-1 and pAAV-INS compared to the cells transfected with pAAV-INS or the cells co-transfected with pAAV-PDX-1 and pAAV-INS.【Conclusion】 The PDX-1 gene affects the expression of the Insulin-secreting cell related genes in 293T cells and plays a potential role in the induction of non-β-cells into the insulin-secreting cells.