MicroRNA-214-3p Regulates Hypoxia-Mediated Pulmonary Artery Smooth Muscle Cell Proliferation and Migration by Targeting ARHGEF12

Medical Science Monitor : International Medical Journal of Experimental and Clinical Research
Xi-Qian XingJiao Yang

Abstract

BACKGROUND miR-214-3p has been found to inhibit proliferation and migration in cancer cells. The objective of this study was to determine whether ARHGEF12 is involved in miR-214-3p-mediated suppression of proliferation and migration of pulmonary artery smooth muscle cells (PASMCs). MATERIAL AND METHODS PASMCs were cultured under normoxia or hypoxia. miR-214-3p mimics or inhibitors were transiently transfected into PASMCs. Proliferation, apoptosis, and migration of PASMCs were evaluated using MTT assay, flow cytometry, and Boyden chamber apparatus. Western blot analysis was used to examine expression of Rho guanine nucleotide exchange factor 12 (ARHGEF12), c-fos, c-jun, and caspase-3. Luciferase reporter assay was used to test the direct regulation of miR-214-3p on the 3'-untranslated region (UTR) of ARHGEF12. RESULTS miR-214-3p was significantly upregulated in hypoxia-treated PASMCs. Knockdown of miR-214-3p significantly attenuated hypoxia-induced proliferation and migration in PASMCs and promoted apoptosis, whereas this effect was aggravated by overexpression of miR-214-3p. In addition, dual-luciferase reporter assay demonstrated that ARHGEF12 is a direct target gene of miR-214-3p. The protein levels of ARHGEF12 were downregul...Continue Reading

Citations

Sep 29, 2020·Hypertension Research : Official Journal of the Japanese Society of Hypertension·Ji-Ru Zhang, Hai-Jian Sun
Sep 19, 2020·Journal of Animal Science and Biotechnology·Shengjie ShiGuiyan Chu
Dec 10, 2020·JACC. Basic to Translational Science·Cátia A Santos-FerreiraHenrique M Girão
Jul 27, 2021·Small GTPases·Neda Z GhanemJoe W Ramos

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Methods Mentioned

BETA
GTPase
nucleotide exchange
ELISA
flow cytometry
Protein Assay
electrophoresis
PCR

Software Mentioned

miRGene
miRbase
SPSS statistics
TargetScan5

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