MiR-449a suppresses proliferation of glomerular mesangial cells cultured in high glucose by inhibiting Trim11

Wenyun CHEN; Yan SUN; Rui PENG; Huimin PENG; Zheng ZHANG

doi:10.16016/j.1000-5404.201903081

您当前的位置：

首页 >

文章列表页 >

MiR-449a suppresses proliferation of glomerular mesangial cells cultured in high glucose by inhibiting Trim11

Basic Medicine | 更新时间：2026-01-09

- MiR-449a suppresses proliferation of glomerular mesangial cells cultured in high glucose by inhibiting Trim11
- Journal of Third Military Medical University Vol. 41, Issue 16, Pages: 1520-1526(2019)
- 作者机构：
  
  1.400016重庆，重庆医科大学基础医学院:细胞生物学及遗传学教研室
  2.400016重庆，重庆医科大学基础医学院:分子肿瘤研究中心
- 作者简介：
- 基金信息：
  
  the Natural Science Foundation of Chongqing(CSTC2018jcyjAX0244)
- DOI：10.16016/j.1000-5404.201903081
  CLC： R322.61;R329.28;R394.1
- Received：10 March 2019，
  
  Revised：2019-04-14，
  
  Published：30 August 2019
- 稿件说明：
移动端阅览
Wenyun CHEN, Yan SUN, Rui PENG, et al. MiR-449a suppresses proliferation of glomerular mesangial cells cultured in high glucose by inhibiting Trim11[J]. Journal of Third Military Medical University, 2019, 41(16): 1520-1526.
DOI：

Wenyun CHEN, Yan SUN, Rui PENG, et al. MiR-449a suppresses proliferation of glomerular mesangial cells cultured in high glucose by inhibiting Trim11[J]. Journal of Third Military Medical University, 2019, 41(16): 1520-1526. DOI： 10.16016/j.1000-5404.201903081.

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(

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(

0.01)；沉默miR-449a，细胞增殖增加，Trim11蛋白水平显著上调(

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.

Methods

RT-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.

Results

Compared with the cells cultured in normal glucose

the mesangial cells exposed to high glucose showed significantly decreased miR-449a expression (

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 (

0.01)

while miR-449a silencing significantly promoted the cell proliferation and increased the expression of Trim11 protein (

0.01). Silencing Trim11 in the cells obviously abolished the proliferation-promoting effect of miR-449a.

Conclusion

MiR-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.

关键词

Keywords

references

XU Y, WANG L, HE J, et al . Prevalence and control of diabetes in Chinese adults[J ] . JAMA, 2013, 310(9): 948-959. DOI: 10.1001/jama. 2013.168118.

ZHANG L X, LONG J Y, JIANG W S, et al . Trends in chronic kidney disease inChina[J ] . N Engl J Med, 2016, 375(9): 905-906. DOI: 10.1056/NEJMc1602469.

XIE Y, BOWE B, MOKDAD A H, et al . Analysis of the Global Burden of Disease study highlights the global, regional, and national trends of chronic kidney disease epidemiology from 1990 to 2016[J ] . Kidney Int, 2018, 94(3): 567-581. DOI: 10.1016/j.kint.2018.04.011.

ZHU M J, CHEN Q, LIU X, et al .LncRNA H19/miR-675 axis represses prostate cancer metastasis by targeting TGFBI[J ] . FEBS J, 2014, 281(16): 3766-3775. DOI: 10.1111/febs.12902.

YAN J, GUO X Q, XIA J Z, et al . MiR-148a regulates MEG3 in gastric cancer by targeting DNAmethyltransferase 1[J ] . Med Oncol, 2014, 31(3): 879. DOI: 10.1007/s12032-014-0879-6.

[HAN Y H, LIU Y C, ZHANG H, et al . Hsa-miR-125b suppresses bladder cancer development by down-regulating oncogene SIRT7 and oncogenic long noncoding RNA MALAT1[J ] . FEBS Lett, 2013.[Epub ahead of print ] . DOI: 10.1016/j.febslet.2013.10.023 http://dx.doi.org/10.1016/j.febslet.2013.10.023 ..

SUN Y, PENG R, PENG H M, et al . MiR-451 suppresses the NF-kappaB-mediated proinflammatory molecules expression through inhibiting LMP7 in diabetic nephropathy[J ] . Mol Cell Endocrinol, 2016, 433: 75-86. DOI: 10.1016/j.mce.2016.06.004.

NOONAN E J, PLACE R F, POOKOT D, et al . MiR-449a targets HDAC-1 and induces growth arrest in prostatecancer[J ] . Oncogene, 2009, 28(14): 1714-1724. DOI: 10.1038/onc.2009.19.

NOONAN E J, PLACE R F, BASAK S, et al . MiR-449a causesRb-dependent cell cycle arrest and senescence in prostate cancer cells[J ] . Oncotarget, 2010, 1(5): 349-358. DOI: 10.18632/oncotarget.167.

LI A L, PENG R, SUN Y, et al .LincRNA 1700020I14Rik alleviates cell proliferation and fibrosis in diabetic nephropathy via miR-34a-5p/Sirt1/HIF-1α signaling[J ] . Cell Death Dis, 2018, 9(5): 461. DOI: 10.1038/s41419-018-0527-8..

ZHANG Z, PENG H M, CHEN J X, et al . MicroRNA-21 protects frommesangial cell proliferation induced by diabetic nephropathy in db/db mice[J ] . FEBS Lett, 2009, 583(12): 2009 -2014. DOI: 10.1016/j.febslet.2009.05.021.

SONG R, WALENTEK P, SPONER N, et al . MiR-34/449miRNAs are required for motile ciliogenesis by repressing cp110[J ] . Nature, 2014, 510(7503): 115-120. DOI: 10.1038/nature13413.

LIU X D, ZHANG L Y, ZHU T C, et al . Overexpression of miR-34c inhibits high glucose-induced apoptosis inpodocytes by targeting Notch signaling pathways[J ] . Int J Clin Exp Pathol, 2015, 8(5): 4525-4534.

ZHANG X Y, SONG S P, LUO H X. Regulation ofpodocyte lesions in diabetic nephropathy via miR-34a in the Notch signaling pathway[J ] . Medicine (Baltimore), 2016, 95(44): e5050.DOI: 10.1097/MD.0000000000005050.

ZHANG L, HE S Y, GUO S D, et al . Down-regulation of miR-34a alleviatesmesangial proliferation in vitro and glomerular hypertrophy in early diabetic nephropathy mice by targeting GAS1[J ] . J Diabetes Complicat, 2014, 28(3): 259-264. DOI: 10.1016/j.jdiacomp.2014.01.002..

SANDBOTHE M, BUURMAN R, REICH N, et al . The microRNA-449 family inhibits TGF-β-mediated liver cancer cell migration by targetingSOX4[J ] . J Hepatol, 2017, 66(5): 1012-1021. DOI: 10.1016/j.jhep.2017.01.004.

YE W W, XUE J S, ZHANG Q, et al . MiR-449a functions as a tumor suppressor in endometrial cancer by targeting CDC25A[J ] . Oncol Rep, 2014, 32(3): 1193-1199. DOI: 10.3892/or.2014.3303.

LIU J J, RAO J, LOU X M, et al . Upregulated TRIM11 exerts its oncogenic effects in hepatocellular carcinoma through inhibition of P53[J ] . Cell Physiol Biochem, 2017, 44(1): 255-266. DOI: 10.1159/000484678.

YIN Y, ZHONG J, LI S W, et al . TRIM11, a direct target of miR-24-3p, promotes cell proliferation and inhibits apoptosis in coloncancer[J ] . Oncotarget, 2016, 7(52): 86755-86765. DOI: 10.18632/oncotarget.13550.

WANG X L, SHI W P, SHI H C, et al . TRIM11 overexpression promotes proliferation, migration and invasion of lung cancer cells[J ] . J Exp Clin Cancer Res, 2016, 35(1): 100. DOI: 10.1186/s13046-016-0379-y.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Promoting effect of ultra-low dose DMSO on fibroblast proliferation and its underlying mechanism

GPX7 regulates proliferation and senescence of hepatocellular carcinoma cells through the PI3K-AKT pathway

Mechanism of abnormal specification of spermatogonial stem cells after Rb1 knockout in mitotic-arrested prospermatogonia

Function of circular RNA hsa_circ_0000711 in hepatocellular carcinoma cell lines

Related Author

TAN Ranjing

KANG Tehao

WU Dengyan

WANG Ruiguan

WANG Jing

YAN Maohui

LI Xinji

SUN Zhijia

Related Institution

Center for Dermatology and Medical Cosmetology， the Second Affiliated Hospital of Chongqing Medical University

Department of Dermatology， Plastic and Cosmetic Surgery， Wushan People’s Hospital

Faculty of Hepatobiliary and Pancreatic Surgery， First Medical Center， General Hospital of PLA， Beijing, Air Force Medical Center of PLA

Department of Hepatobiliary Surgery， Eighth Medical Center， General Hospital of PLA， Beijing, Air Force Medical Center of PLA

Department of Radiation Oncology, Air Force Medical Center of PLA

⁰