miR-216a and miR-301a promote the proliferation of pulmonary arterial smooth muscle cells under hypoxia via regulating BMPR2
LIN Xiao1, ZHENG Weiping1, LI Hongru2, CHEN Yusheng2, ZHOU Meng3
（1. Department of Geriatric Medicine, 2. Department of Respiratory and Critical Care Medicine, 3. Department of Rheumatology and Immunology, Fujian Provincial Hospital, Fuzhou Fujian 350001, China）
Abstract：Objective: To investigate the effects of miR-216a and miR-301a on the proliferation of human pulmonary arterial smooth muscle cells （HPASMC） under hypoxia and to explore the potential mechanisms. Methods: The proliferation activity of HPASMC under normoxia and different hypoxia time was detected by CCK8 assay, and qRT-PCR was used to detect the expression of miR-216a, miR-301a and bone morphogenetic protein receptor type 2 （BMPR2） mRNA in each group. The proliferation ability and proliferating cell nuclear antigen （PCNA） expression levels of HPASMC transfected with anti-miR-216a or anti-miR-301a mimics were detected by CCK8 assay and Western blotting, respectively. The relationship between BMPR2 and miR216a, BMPR2 and miR301a were detected by dual luciferase reporter assay. CCK8 assay and Western blot were used to detect the proliferation ability of HPASMC and the expression levels of BMPR2 and PCNA after simultaneous transfection of miR-216a or miR-301a combined with BMPR2 overexpression plasmid, respectively. Results: With the prolongation of hypoxia time, the proliferation activity of HPASMC and the expression of miR-216a increased, while the expression of BMPR2 mRNA gradually decreased. The expression level of miR-301a was the highest in HPASMC exposed to hypoxia for 24 hours. Down-regulation of miR-216a and miR-301a significantly inhibited the proliferation activity and PCNA expression of HPASMC. Dual luciferase reporter assay confirmed that BMPR2 was a target of miR-216a and miR-301a, respectively. Simultaneously transfected with miR-216a or miR-301a combined with BMPR2 overexpression plasmid, the expression of BMPR2 decreased, and the cell proliferation activity and PCNA expression increased. Conclusion: miR-216a and miR-301a induce pulmonary arterial hypertension by promoting the proliferation of HPASMC under hypoxia via inhibiting the expression of BMPR2.
［Key words］miR-216a; miR-301a; pulmonary arterial hypertension; pulmonary arterial smooth muscle cells; bone morphogenetic protein receptor type 2
