FTO过表达对小鼠胰岛β细胞功能的影响以及全基因组表达谱变化研究

Effect of FTOoverexpression on mouse pancreatic islet β cells function and differential gene expression profile

为了研究肥胖基因FTO过表达对小鼠胰岛β细胞功能的影响以及基因表达谱的变化,构建小鼠FTO基因过表达慢病毒载体,包装慢病毒颗粒并感染小鼠胰岛MIN6细胞。利用QPCR和Western Blot技术检测FTO基因在MIN6细胞中过表达情况,并检测葡萄糖刺激检测胰岛素的释放情况,进一步利用小鼠全基因芯片检测FTO过表达对小鼠胰岛MIN6细胞表达谱的影响。结果表明:慢病毒载体成功介导了FTO在MIN6细胞中过表达,FTO过表达可以显著抑制MIN6细胞的胰岛素释放。表达芯片的结果显示FTO改变MIN6细胞表达谱,发现多达922个的差异基因。FTO过表达改变了小鼠胰岛细胞的表达谱,差异基因通过一些重要信号通路影响小鼠胰岛β细胞的生物学功能。

To explore the effect of FTO (fat mass and obesity associated)overexpression on mouse pancreatic islet βcells and genes expression profile,mouse FTO gene was cloned into the lentivirus.Lentivirus was packaged in 293T cells and delivered in-to the pancreatic islet cells.QPCR and Western blot assay were employed to detect the overexpression of FTO in MIN6 cells. Glucose-stimulated insulin secretion (GSIS)was performed to detect the insulin secretion of MIN6 cells.Mouse gene microarray was used to detect the differential gene expression profile of the islet cells modulated by FTO overexpression.MIN6 cells stably expressing FTO was achieved by lentivirus delivery.FTO overexpression significantly inhibits the insulin secretion.The microar-ray shows that there are totally 922 differential expression genes.FTO overexpression changes the gene expression profile of MIN6 cells.The differentially expressed genes may regulate the isletβcells functions via important signal pathways.