Effects of miR‑195‑5p on cell proliferation and apoptosis in gestational diabetes mellitus via targeting EZH2

Abstract

Gestational diabetes mellitus (GdM) is a type of diabetes mellitus (dM) that occurs during pregnancy. The present study aimed to investigate the roles of microrna (mir)-195-5p and enhancer of zeste homolog 2 (eZH2) in GdM, and their potential association. Human umbilical vein endothelial cells (HuVecs) were collected from healthy and GdM umbilical cords, and the endothelial properties were detected by flow cytometry. mRNA expression levels of miR-195-5p and EZH2, and EZH2 protein expression levels were detected by reverse transcription-quantitative Pcr (rT-qPcr) and western blot analysis, respectively. cell colony formation and flow cytometry were performed to determine cell proliferation and apoptosis. Furthermore, the target gene of mir-195-5p was predicted and assessed using a dual-luciferase reporter assay. The levels of cell viability, proliferation and apoptosis following the overexpression of miR-195-5p, EZH2 or mir-195-5p + eZH2, were detected using cell counting Kit-8, colony formation and flow cytometry assays, respectively. In addition, the mRNA expression levels of miR-195-59 and EZH2, and EZH2 protein expression levels following transfection with overexpression plasmids were detected using rT-qPcr and western blot analysis, respectively. it was identified that high mRNA expression of miR-195-5p, and low EZH2 mRNA and protein expression levels decreased the level of cell proliferation and the high apoptotic rate of GdM-HuVecs. in addition, mir-195-5p was predicted and identified to target EZH2, and miR-195-5p overexpression was identified to inhibit cell proliferation and promote apoptosis. However, it was demonstrated that upregulation of eZH2 could alleviate the inhibition of cell proliferation and the increased apoptotic rate induced by miR-195-5p overexpression. Therefore, the present results suggested that mir-195-5p may inhibit cell viability, proliferation and promote apoptosis by targeting eZH2 in GdM-induced HuVecs.

Epigenetics, microRNA and Metabolic Syndrome: A Comprehensive Review

“…Expression of miR-101-3p, a pancreatic islet-enriched miRNA with a role in insulin secretion and β cell functioning [ 85 ], is demonstrated to be regulated by H3K27me3 modification in EZH2. Moreover, epigenetically regulated alterations in miRNAs induced by suboptimal maternal nutrition or endocrine factors are also reported to be responsible for altered gene expression and to promote offspring and adult MetS phenotypes [ 86 , 87 ]. Altogether, these studies provide evidence for a functional link between classical epigenetic regulations of miRNA underpinning pathophysiological processes characterised as causal factors in the development of MetS.…”

The Endothelium as a Target for Anti-Atherogenic Therapy: A Focus on the Epigenetic Enzymes EZH2 and SIRT1

“…The molecular mechanisms by which EZH2 may affect endothelial-dependent atherogenesis are gaining attention. The independent atherosclerosis risk factors dyslipidaemia [ 130 ], hyperhomocysteinemia, [ 144 ] hyperglycaemia [ 156 ] and OSS [ 110 ] all block the expression of endothelial microRNAs that repress EZH2 translation under physiological conditions. For example, in human umbilical vein endothelial cells (HUVECs) treated with oxLDL, miR-200a expression is decreased, causing an upregulation of EZH2 [ 130 ].…”

“…For example, in human umbilical vein endothelial cells (HUVECs) treated with oxLDL, miR-200a expression is decreased, causing an upregulation of EZH2 [ 130 ]. During hyperhomocysteinemia, levels of EZH2 are increased as a result of reduced levels of miR-92a [ 144 ] and upon OSS, miR-101 expression is downregulated [ 110 , 157 ] also leading to an increased expression of endothelial EZH2 and the concurrent H3K27Me3 [ 130 , 144 , 156 ]. Interestingly, in the field of oncology, multiple other microRNAs such as miR-26a, miR-124 and miR-214 have been described to repress EZH2 expression [ 158 , 159 ].…”

“…Next, long non-coding RNAs (lncRNAs) guide EZH2 towards target genes [ 120 ], where critical endothelial genes are silenced. This orchestrated mechanism ultimately culminates in endothelial inflammatory activation, dysfunction and cell death [ 120 , 130 , 144 , 156 ].…”

Non-Coding RNAs and Extracellular Vehicles: Their Role in the Pathogenesis of Gestational Diabetes Mellitus

“…They found that 11 miRNAs (including hsa-miR-1299, hsa-miR-1200, hsa-miR-578, hsa-miR-593-5p, hsa-miR-765, hsa-miR-520d-3p, hsa-miR-617, hsa-miR-92a-3p, hsa-miR-30b-5p, hsa-miR-181c-5p, and hsa-miR-181a-5p) differed in their expression ( 99 ), indicating their potential role in GDM; however, future studies are needed to elucidate this further. Additionally, miR-195-5p was up-regulated in GDM-induced HUVECs, inhibited cell viability and proliferation, and promoted apoptosis by targeting EZH2 in vitro ( 72 ), thus highlighting the importance of miRNAs in GDM.…”