Department of Hepatobiliary Surgery, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China.
Department of Vascular Surgery, Qianwei Hospital of Jilin Province, Changchun, Jilin 130012, P.R. China.
Mol Med Rep. 2021 Sep;24(3). doi: 10.3892/mmr.2021.12266. Epub 2021 Jul 2.
Varicose veins are among the most common disorders of the vascular system; however, the pathogenesis of varicose veins remains unclear. The present study aimed to investigate the roles of microRNA (miR)‑199a‑5p in varicose veins and in the phenotypic transition of vascular smooth muscle cells (VSMCs). Bioinformatics analysis confirmed that miR‑199a‑5p had target sites on the forkhead box C2 (FOXC2) 3'‑untranslated region. Reverse transcription‑quantitative PCR (RT‑qPCR) and western blotting were used to detect the expression levels of miR‑199a‑5p and FOXC2 in varicose vein and normal great saphenous vein tissues. Cell Counting Kit‑8 and Transwell migration assays were performed to validate the effects of miR‑199a‑5p on VSMCs. Contractile markers, such as smooth muscle 22α, calponin, smooth muscle actin and myosin heavy chain 11 were used to detect phenotypic transition. RT‑qPCR revealed that miR‑199a‑5p was downregulated in varicose veins compared with expression in normal great saphenous veins, whereas FOXC2 was upregulated in varicose veins. In addition, biomarkers of the VSMC contractile phenotype were downregulated in varicose veins. Overexpression of miR‑199a‑5p by mimics suppressed VSMC proliferation and migration, whereas depletion of miR‑199a‑5p enhanced VSMC proliferation and migration. Notably, the effects caused by miR‑199a‑5p could be reversed by FOXC2 overexpression. Dual luciferase reporter analysis confirmed that FOXC2 was a target of miR‑199a‑5p. In conclusion, miR‑199a‑5p may be a novel regulator of phenotypic switching in VSMCs by targeting FOXC2 during varicose vein formation.
静脉曲张是血管系统中最常见的疾病之一;然而,静脉曲张的发病机制仍不清楚。本研究旨在探讨微小 RNA(miR)-199a-5p 在静脉曲张和血管平滑肌细胞(VSMC)表型转化中的作用。生物信息学分析证实 miR-199a-5p 在叉头框 C2(FOXC2)3'非翻译区有靶位。逆转录-定量聚合酶链反应(RT-qPCR)和 Western blot 用于检测 miR-199a-5p 和 FOXC2 在静脉曲张和正常大隐静脉组织中的表达水平。细胞计数试剂盒-8 和 Transwell 迁移实验用于验证 miR-199a-5p 对 VSMC 的影响。收缩标志物,如平滑肌 22α、钙调蛋白、平滑肌肌动蛋白和肌球蛋白重链 11,用于检测表型转化。RT-qPCR 显示,与正常大隐静脉相比,miR-199a-5p 在静脉曲张中表达下调,而 FOXC2 在静脉曲张中表达上调。此外,静脉曲张中 VSMC 收缩表型的生物标志物表达下调。模拟物过表达 miR-199a-5p 抑制 VSMC 增殖和迁移,而 miR-199a-5p 耗竭增强 VSMC 增殖和迁移。值得注意的是,miR-199a-5p 的作用可被 FOXC2 过表达逆转。双荧光素酶报告基因分析证实 FOXC2 是 miR-199a-5p 的靶标。综上所述,miR-199a-5p 可能通过靶向 FOXC2 调节 VSMC 表型转换,从而成为静脉曲张形成过程中的一种新的调控因子。