沉默PMR1表达促进胰岛细胞分泌胰岛素的研究  

Plasma membrane-related Ca^(2+)-ATPase-1 gene silencing promotes insulin secretion in islet β cells NIT

作　　者：房彦平[1] 季爱民[2] 杨月莲[1] 冯巧飞[1] 孙靓[2] 刘宏[1] 

机构地区：[1]南方医科大学珠江医院内分泌科,广东广州510282 [2]南方医科大学珠江医院药剂科,广东广州510282

出　　处：《南方医科大学学报》2009年第8期1565-1567,1570,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30640053)

摘　　要：目的利用RNA干扰技术(RNAi)研究体外合成的siRNA分子对高尔基体膜上钙离子ATP酶1(plasma membrane-relatedCa2+-ATPase-1,PMR1)的基因沉默作用,并探讨沉默后对胰岛细胞分泌胰岛素的影响。方法委托公司合成抗PMR1基因的siPMR1,用脂质体方法介导siPMR1转染胰岛NIT-1细胞,采用RT-PCR法观察siPMR1转染前后胰岛细胞中PMR1mRNA的表达变化,并用放射免疫分析法测定胰岛细胞分泌的胰岛素浓度。结果siPMR1转染胰岛NIT-1细胞后能明显抑制PMR1mRNA表达,siPMR1转染的胰岛NIT-1细胞能显著增加胰岛素分泌量。结论siPMR1可明显沉默PMR1基因表达,并促进胰岛细胞分泌胰岛素,为用RNAi技术沉默PMR1靶点治疗糖尿病提供了理论基础。Objective To assess the effect of RNA interference-mediated gene silencing of plasma membrane-related Ca2+-ATPase-1 (PMR1) gene on the insulin secretion in islet β cells NIT-1 in vitro. Methods A small interfering RNA duplex (siPMR1) corresponding to the nucleotides 337-357 of mouse PMR1 cDNA was introduced into NIT-1 cells via liposomes. The gene silencing effect was assessed by RT-PCR, and the total insulin level in the transfected cells was measured by radioimmunoassay. Results Transfection with siPMR1 r...

关 键 词：膜上钙离子ATP酶1 SIRNA RNA干扰技术 胰岛素 

分 类 号：Q789[生物学—分子生物学]