LncRNA HOTTIP通过调控miR-101对胰岛β细胞功能的影响

Effects of lncRNA HOTTIP on isletβcell function by regulating miR-101

目的探讨lncRNA HOTTIP对胰岛β细胞功能的作用及潜在机制。方法选取30例GDM孕妇为GDM组,以及30例健康孕妇为对照组。培养大鼠胰岛素瘤细胞系INS-1细胞,将Vector、pcDNA-HOTTIP、NC-siRNA、HOTTIP-siRNA、pcDNA-HOTTIP+NC mimic、pcDNA-HOTTIP+miR-101 mimic分别转染至INS-1细胞,RT-qPCR实验检测lncRNA HOTTIP、miR-101的表达水平;葡萄糖测定试剂盒检测血糖水平;ELISA方法测量胰岛素分泌水平;CCK-8方法测定细胞增殖能力;生物信息学软件和双荧光素酶报告基因实验预测和验证lncRNA HOTTIP和miR-101之间的靶向关系。结果GDM孕妇中血糖水平升高,lncRNA HOTTIP表达水平下调,miR-101表达水平显著上调(P<0.05)。过表达HOTTIP促进INS-1细胞增殖能力和胰岛素分泌水平(P<0.05);敲低HOTTIP抑制INS-1细胞增殖能力和胰岛素分泌水平(P<0.05)。LncRNA HOTTIP直接靶向miR-101并负调控其表达,过表达miR-101可显著逆转HOTTIP对胰岛β细胞功能的作用。结论lncRNA HOTTIP过表达可通过调控miR-101促进INS-1细胞增殖和胰岛素产生。

Objective To investigate the effect of lncRNA HOTTIP on isletβcell function and its potential mechanism.Methods Thirty pregnant women with GDM were selected as GDM group and 30 healthy pregnant women as control group.INS-1 cells of rat insulinoma cell line were cultured.Vector,pcDNA-HOTTIP,NC-SiRNA,HOTTIP-siRNA,pcDNA-HOTTIP+NC mimic,and pcDNA-HOTTIP+miR-101 mimic were transfected into INS-1 cells,respectively.The levels of HOTTIP and miR-101 of lncRNA were detected by RT-qPCR.Glucose assay kit was used to detect blood glucose level.Insulin secretion was measured by ELISA.Cell proliferation was measured by CCK-8 method.Bioinformatics software and dual luciferase reporter gene experiments predicted and verified the targeting relationship between lncRNA HOTTIP and miR-101.Results In pregnant women with GDM,blood glucose level was increased,level of lncRNA HOTTIP was down-regulated,and level of miR-101 was significantly up-regulated(P<0.05).Overexpression of HOTTIP promoted the proliferation and insulin secretion of INS-1 cells(P<0.05).Knocking down HOTTIP inhibited proliferation of INS-1 cell and insulin secretion(P<0.05).LncRNA HOTTIP directly targets miR-101 and negatively regulates its expression.Overexpression of miR-101 can significantly reverse the effect of HOTTIP on the function of isletβcells.Conclusion lncRNA HOTTIP overexpression can promote INS-1 cell proliferation and insulin production by regulating miR-101.