Mir-27A-3P Inhibits Type I And Iii Collagen Synthesis In Lung Fibroblasts Through The Wnt3A/¦¢-Catenin Signaling Pathway
Keywords
Mir-27A-3P, Wnt3A, ¦¢-Catenin, Type ¢ñ Collagen, Type ¢ó Collagen, Pulmonary Fibrosis
Abstract
Objective To explore the effect and molecular mechanism of microRNA-27a-3p (miR-27a-3p) on the synthesis of type I collagen (Col ¢ñ) and type III collagen (Col ¢ó) in lung fibroblasts. Methods Human embryonic lung fibroblast cells MRC-5 were cultured, transfected with miR-27a-3p mimic/inhibitor, and real-time fluorescence quantitative PCR (qPCR) was used to detect the level of miR-27a-3p; qPCR and Western blot were used to determine Col ¢ñ and Col ¢ó. , Wnt3a expression; Western blot analysis of ¦Â-catenin levels in the nucleus. Bioinformatics was used to predict the targeted binding of miR-27a-3p to Wnt3a 3′-untranslated region (3′-UTR), and a dual-luciferase reporter gene was used to detect the effect of miR-27a-3p mimics on Wnt3a 3′-UTR wild Effects of type and mutant luciferase activity. MRC-5 cells were pretreated with Wnt3a/¦Â-catenin signaling pathway inhibitor Dkk1 for 6 h, and then transfected with miR-27a-3p inhibitor for 48 h. The expression of Col ¢ñ and Col ¢ó was detected by qPCR and Western blot. Results: The miR-27a-3p mimic significantly increased the level of miR-27a-3p and reduced the expression of Col ¢ñ, Col ¢ó, Wnt3a and ¦Â-catenin. The effect of its inhibitor was opposite. Compared with the control group, the differences were significant ( P<0.01). There is a binding site for miR-27a-3p in Wnt3a 3'-UTR. Overexpression of miR-27a-3p reduced wild-type Wnt3a 3'-UTR luciferase activity (P<0.01), but not mutant luciferase. Enzyme activity had no effect (P>0.05). Dkk1 pretreatment almost completely reversed the induction effect of miR-27a-3p inhibitor on the synthesis of Col ¢ñ and Col ¢ó (P<0.05). Conclusion miR-27a-3p inhibits the biosynthesis of Col ¢ñ and Col ¢ó in lung fibroblasts, and its mechanism is related to antagonizing the Wnt3a/¦Â-catenin signaling pathway. For further information of this article and research, feel free to contact our team for asssitance. Original research was done by 10.3969/j.issn.1001-1978.2019.02.017
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