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

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/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 downregulated after knockdown of miR-214-3p in PASMCs. Rescue experiment results indicated that decreased proliferation of PASMCs resulted from knockdown of miR-214-3p were partially reversed by silencing of ARHGEF12 by siRNA. Furthermore, knockdown of miR-214-3p reduced expression of c-jun and c-fos, but increased expression of caspase-3 in PASMCs under hypoxia. Conclusions In conclusion, these results indicate that miR-214-3p acts as a novel regulator of hypoxia-induced proliferation and migration by directly targeting ARHGEF12 and dysregulating c-jun and c-fos in PASMCs, and may be a potential therapeutic target for treating pulmonary hypertension.

MiR-214-3p Promotes Proliferation and Inhibits Estradiol Synthesis In Porcine Granulosa Cells

“…2D-F and 3D-F). Cyclin B is a marker of immunohistochemical proliferation (37) and CDK4 is a kinase that regulates the transition from the G1 to S phases of the cell cycle (39). We found that due to miR-214-3p agomir and antagomir, compared to our NC, Cyclin B and CDK4 had the most significant differential expression of mRNA and protein levels.…”

“…Previous research has shown that miR-214-3p regulates the proliferation of breast cancer cells by targeting survivin protein (36) and can promote smooth muscle cell proliferation (37), which indicates that miR-214-3p is indeed involved in cell proliferation processes. We transfected miR-214-3p agomir and antagomir into porcine ovarian GCs in this study to explore the effects of miR-214-3p on GCs proliferation.…”

Therapeutic Benefit of the Association of Lodenafil with Mesenchymal Stem Cells on Hypoxia-induced Pulmonary Hypertension in Rats

“…A downstream signaling molecule related to the proliferation phenotype in PAH is c-fos, whose expression is increased at pulmonary arterioles and is responsible for enhanced pulmonary artery smooth muscle cell proliferation in an animal model and in PAH patients [ 35 ]. Since c-fos proliferation in pulmonary artery smooth muscle exposed to hypoxia depended on miRNA-214-3p [ 36 ], hMSCs could regulate local miRNA to promote an anti-proliferative effect. It is possible that miRNA-214-3p could be downregulated by lodenafil + hMSCs because of the reversal of increased c-fos expression in pulmonary arterioles.…”

MiR-214-3p promotes proliferation and inhibits estradiol synthesis in porcine granulosa cells

“…3), which is consistent with previous research results. For example, miR-214-3p regulates the proliferation of breast cancer cells by targeting survivin protein [36] and can promote smooth muscle cell proliferation [37],…”