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30 July 2019 : Laboratory Research  

MicroRNA-154 Targets the Wnt/β-Catenin Signaling Pathway Following Injury to Human Vascular Endothelial Cells by Hydrogen Peroxide

Yan Li1ABCE, Ranran Meng2ABCE*

DOI: 10.12659/MSM.915263

Med Sci Monit 2019; 25:5648-5656

Abstract

BACKGROUND: Endothelial cells are involved in vascular homeostasis, and endothelial cell dysfunction is involved in the pathogenesis of cardiovascular disease. This study aimed to investigate the effects of microRNA-154 in human umbilical vein endothelial cells (HUVECs) following injury induced by hydrogen peroxide (H₂O₂).

MATERIAL AND METHODS: Cell viability and apoptosis of HUVECs treated with H₂O₂ were measured. The expression of microRNA-154 was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell survival, caspase-3 activity, and the apoptosis rate were evaluated in H₂O₂-treated HUVECs cells after the upregulation and down-regulation of microRNA-154 expression. The interaction between microRNA-154 and Dickkopf WNT signaling pathway inhibitor 2 (DKK2) was predicted by bioinformatics analysis and was verified by luciferase reporter gene assay and Western blot. The effects of DKK2 short-interfering RNA (siRNA) on antioxidant injury in HUVECs cells were determined.

RESULTS: The survival rate of HUVECs exposed to H₂O₂ was significantly reduced and the apoptosis rate was significantly increased, and H₂O₂ significantly inhibited the expression of microRNA-154 in a dose-dependent manner. Overexpression of microRNA-154 increased cell survival, reduced the activity of caspase-3, and reduced cell apoptosis. Inhibition of microRNA-154 expression decreased cell survival, increased the activity of caspase-3, and promoted cell apoptosis. Luciferase reporter gene assay and Western blot showed that microRNA-154 interacted with the Wnt pathway molecule DKK2 in HUVECS. Also, DDK2 siRNA resulted in a similar protective effect on H₂O₂-treated HUVECs as overexpression of microRNA-154.

CONCLUSIONS: Oxidative injury in HUVECs was regulated by microRNA-154 targeting the Wnt/β-catenin signaling pathway.

Keywords: Cardiovascular Diseases, Cardiovascular System, endothelial cells, MicroRNAs, Apoptosis, Apoptosis Regulatory Proteins, Caspase 3, Cell Survival, Human Umbilical Vein Endothelial Cells, Hydrogen Peroxide, Intercellular Signaling Peptides and Proteins, Oxidative Stress, Reactive Oxygen Species, Wnt Signaling Pathway

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750