MiR-96靶向调控Sox6促进小鼠β细胞胰岛素合成与分泌  

MiR-96 promotes insulin biosynthesis and secretion in mouse β cells through targeting Sox6

作　　者：杨靖[1] 吴丽 周灵芝[2] 赵志波[1] 项孙敏 颜斌[1] 吕运成[3] 肖新华[1] YANG Jing;WU Li;ZHOU Lingzhi;ZHAO Zhibo;XIANG Sunmin;YAN Bin;Lü Yuncheng;XIAO Xinhua(Department of Metabolism & Endocrinology,the First Affiliated Hospital of the University of South China,Hengyang 421001,China)

机构地区：[1]南华大学附属第一医院内分泌科,湖南衡阳421001 [2]南华大学附属第一医院儿科,湖南衡阳421001 [3]南华大学医学院应用解剖与生殖医学研究所,湖南衡阳421001

出　　处：《实用医学杂志》2019年第2期196-200,共5页The Journal of Practical Medicine

基　　金：湖南省卫计委重大专项(编号:2017011)

摘　　要：目的探索miR-96对小鼠胰岛β细胞胰岛素合成与分泌的影响及其作用机制。方法将小鼠胰岛β细胞系(MIN6细胞)分为四组,通过脂质体LipfectamineTM2000介导将miR-96 mimics、mimicsNC、miR-96 inhibitor、inhibitor NC瞬时转染至4组细胞,荧光定量PCR和ELISA方法分别检测Ins1、Ins2mRNA的表达及细胞上清液中的胰岛素浓度,生物信息学分析miR-96与Sox6 3UTR的结合关系,荧光素酶报告基因检测miR-96与Sox6的结合情况,Western blot检测Sox6蛋白水平。结果与对照组比较,miR-96 mimics能增加葡萄糖刺激下MIN6细胞胰岛素的合成与分泌(P <0.01),而miR-96 inhibitor可下调胰岛素的合成与分泌(P <0.01)。miR-96与Sox6 3UTR的553-561位点结合。过表达miR-96可显著抑制Sox6荧光素酶活性及MIN6细胞中Sox6蛋白的表达。结论 miR-96靶向沉默Sox6进而促进β细胞胰岛素的合成与分泌。Objective To observe the effect of miR-96 on insulin biosynthesis and secretion and to explore the underlying mechanism in mouse isletβcells.Methods Mouse isletβcell line(MIN6 cells)were divided into 4 groups and miR-96 mimics,mimics NC,miR-96 inhibitor,inhibitor NC were transfected into the 4groups of cells,respectively.The levels of Ins1 and Ins2 mRNA in each group of MIN6 cells were detected by realtime PCR,and the insulin secretion was measured by ELISA.The binding site between miR-96 and Sox6 3‘UTR was predicted by bioinformatics analysis and further confirmed by luciferase reporter assay,and the level of Sox6protein was detected by Western blot.ResultsCompared with the control group,miR-96 mimics increased the synthesis and secretion of insulin in MIN6 cells stimulated by glucose(P<0.01),while miR-96 inhibitor downregulated the synthesis and secretion of insulin(P<0.01).miR-96 binds to the 553-561 sites within the 3’UTR of Sox6.miR-96 inhibited the luciferase activity and expression of Sox6 protein in MIN6 cells significantly.Conclusion miR-96 promotes insulin biosynthesis and secretion in mouseβ-cells through targeting Sox6.

关 键 词：miR-96 Sox6基因 胰岛素 胰岛Β细胞 

分 类 号：R587.1[医药卫生—内分泌]