[1]陈雯韵,孙艳,彭睿,等.miR-449a通过Trim11影响高糖培养的肾小球系膜细胞增殖[J].第三军医大学学报,2019,41(16):1520-1526.
 CHEN Wenyun,SUN Yan,PENG Rui,et al.MiR-449a suppresses proliferation of glomerular mesangial cells cultured in high glucose by inhibiting Trim11[J].J Third Mil Med Univ,2019,41(16):1520-1526.
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miR-449a通过Trim11影响高糖培养的肾小球系膜细胞增殖(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第16期
页码:
1520-1526
栏目:
基础医学
出版日期:
2019-08-30

文章信息/Info

Title:
MiR-449a suppresses proliferation of glomerular mesangial cells cultured in high glucose by inhibiting Trim11
作者:
陈雯韵孙艳彭睿彭惠民张政
重庆医科大学基础医学院:细胞生物学及遗传学教研室,分子肿瘤研究中心
Author(s):
CHEN Wenyun SUN Yan PENG Rui PENG Huimin ZHANG Zheng

Department of Cellular Biology and Genetics, Molecular Medicine and Cancer Research Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing, 400016, China

关键词:
肾小球系膜细胞miR-449aTrim11细胞增殖
Keywords:
glomerular mesangial cells miR-449a Trim11 cell proliferation
分类号:
R322.61;R329.28;R394.1
文献标志码:
A
摘要:

目的 探讨在高糖培养的系膜细胞中miR-449a与Trim11结合对细胞增殖的调控作用。方法 RT-qPCR检测miR-449a在高糖(25 mmol/L)和低糖(5.5 mmol/L)培养的系膜细胞中的表达;EdU检测miR-449a及Trim11对系膜细胞增殖的影响;生物信息学预测miR-449a与Trim11的结合位点;双荧光素酶验证miR-449a与Trim11靶向作用;Western blot检测上调、下调miR-449a后Trim11蛋白的变化。结果与低糖组比较,miR-449a在高糖培养的系膜细胞中的表达显著降低(P<0.01);双荧光素酶证实miR-449a直接靶向Trim11,过表达miR-449a后,细胞增殖减少,Trim11蛋白水平显著降低(P<0.01);沉默miR-449a,细胞增殖增加,Trim11蛋白水平显著上调(P<0.01);沉默Trim11后,抑制miR-449a对于细胞增殖促进作用被阻止。结论 miR-449a可抑制系膜细胞增殖,其机制可能与抑制靶基因Trim11的表达有关,可能是糖尿病肾病的重要保护性因子。

Abstract:

Objective To investigate the role of miR-449a and its target gene Trim11 in regulating the proliferation of glomerular mesangial cells cultured in high glucose. MethodsRT-qPCR was used to detect the expression of miR-449a in glomerular mesangial cells cultured in high (25 mmol/L) and normal (5.5 mmol/L) glucose. EdU incorporation assay was used to evaluate the effect of overexpression and inhibition of miR-449a and Trim11 on the proliferation of the mesangial cells. Bioinformatics analysis was used for predicting the binding sites between miR-449a and Trim11, and dual luciferase assay was performed to validate their binding. Western blotting was used to detect the changes in Trim11 protein expression in the cells transfected with miR-449a mimics or its inhibitor. ResultsCompared with the cells cultured in normal glucose, the mesangial cells exposed to high glucose showed significantly decreased miR-449a expression (P<0.01). Dual luciferase assay confirmed Trim11 as the direct target of miR-449a. Overexpression of miR-449a in the mesangial cells significantly suppressed the cell proliferation and lowered the expression of Trim11 protein (P<0.01), while miR-449a silencing significantly promoted the cell proliferation and increased the expression of Trim11 protein (P<0.01). Silencing Trim11 in the cells obviously abolished the proliferation-promoting effect of miR-449a. ConclusionMiR-449a can inhibit the proliferation of glomerular mesangial cells cultured in high glucose possibly through inhibition of its target gene Trim11, suggesting the role of miR-449a as an important protective factor against diabetic nephropathy.

更新日期/Last Update: 2019-08-22