MicroRNA‑199a‑5p regulates FOXC2 to control human vascular smooth muscle cell phenotypic switch

Cao,Yushi; Cao,Yushi; Cao,Zhongwen; Cao,Zhongwen; Wang,Weitie; Wang,Weitie; Jie,Xiangyu; Jie,Xiangyu; Li,Lei; Li,Lei

doi:10.3892/mmr.2021.12266

MicroRNA‑199a‑5p regulates FOXC2 to control human vascular smooth muscle cell phenotypic switch

Authors:
- Yushi Cao
- Zhongwen Cao
- Weitie Wang
- Xiangyu Jie
- Lei Li
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Affiliations: 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, Department of Cardiovascular Surgery, The Second Hospital of Jilin University, Changchun, Jilin 130041, P.R. China
Published online on: July 1, 2021 https://doi.org/10.3892/mmr.2021.12266
Article Number: 627
Copyright: © Cao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

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.

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1	Piazza G: Varicose veins. Circulation. 130:582–587. 2014. View Article : Google Scholar : PubMed/NCBI
2	Jacobs BN, Andraska EA, Obi AT and Wakefield TW: Pathophysiology of varicose veins. J Vasc Surg Venous Lymphat Disord. 5:460–467. 2017. View Article : Google Scholar : PubMed/NCBI
3	Xu Y, Bei Y, Li Y and Chu H: Phenotypic and functional transformation in smooth muscle cells derived from varicose veins. J Vasc Surg Venous Lymphat Disord. 5:723–733. 2017. View Article : Google Scholar : PubMed/NCBI
4	Huang X, Liu Z, Shen L, Jin Y, Xu G, Zhang Z, Fang C, Guan W and Liu C: Augmentation of miR-202 in varicose veins modulates phenotypic transition of vascular smooth muscle cells by targeting proliferator-activated receptor-γ coactivator-1α. J Cell Biochem. 120:10031–10042. 2019. View Article : Google Scholar : PubMed/NCBI
5	Qi XB, Jia B, Wang W, Xu GH, Guo JC, Li X and Liu JN: Role of miR-199a-5p in osteoblast differentiation by targeting TET2. Gene. 726:1441932020. View Article : Google Scholar : PubMed/NCBI
6	Bao N, Fang B, Lv H, Jiang Y, Chen F, Wang Z and Ma H: Upregulation of miR-199a-5p protects spinal cord against ischemia/reperfusion-induced injury via downregulation of ECE1 in rat. Cell Mol Neurobiol. 38:1293–1303. 2018. View Article : Google Scholar : PubMed/NCBI
7	Zhu H: Forkhead box transcription factors in embryonic heart development and congenital heart disease. Life Sci. 144:194–201. 2016. View Article : Google Scholar : PubMed/NCBI
8	Liu Y, Liu G, Zhang H and Wang J: MiRNA-199a-5p influences pulmonary artery hypertension via downregulating Smad3. Biochem Biophys Res Commun. 473:859–866. 2016. View Article : Google Scholar : PubMed/NCBI
9	Huang Y, Zhang S, Fang X, Qin L, Fan Y, Ding D, Liu X and Xie M: Plasma miR-199a-5p is increased in neutrophilic phenotype asthma patients and negatively correlated with pulmonary function. PLoS One. 13:e01935022018. View Article : Google Scholar : PubMed/NCBI
10	Agarwal V, Bell GW, Nam JW and Bartel DP: Predicting effective microRNA target sites in mammalian mRNAs. Elife. 4:e050052015. View Article : Google Scholar : PubMed/NCBI
11	Mellor RH, Brice G, Stanton AW, French J, Smith A, Jeffery S, Levick JR, Burnand KG and Mortimer PS; Lymphoedema Research Consortium, : Mutations in FOXC2 are strongly associated with primary valve failure in veins of the lower limb. Circulation. 115:1912–1920. 2007. View Article : Google Scholar : PubMed/NCBI
12	Guo Z, Luo C, Zhu T, Li L and Zhang W: Elevated c-fos expression is correlated with phenotypic switching of human vascular smooth muscle cells derived from lower limb venous varicosities. J Vasc Surg Venous Lymphat Disord. 9:242–251. 2021. View Article : Google Scholar : PubMed/NCBI
13	Surendran S, Ramegowda KS, Suresh A, Binil Raj SS, Lakkappa RK, Kamalapurkar G, Radhakrishnan N and C Kartha C: Arterialization and anomalous vein wall remodeling in varicose veins is associated with upregulated FoxC2-Dll4 pathway. Lab Invest. 96:399–408. 2016. View Article : Google Scholar : PubMed/NCBI
14	Baeten JT and Lilly B: Notch signaling in vascular smooth muscle cells. Adv Pharmacol. 78:351–382. 2017. View Article : Google Scholar : PubMed/NCBI
15	Porter JM and Moneta GL: Reporting standards in venous disease: An update. International Consensus Committee on Chronic Venous Disease. J Vasc Surg. 21:635–645. 1995. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
17	Tang Y, Yu S, Liu Y, Zhang J, Han L and Xu Z: MicroRNA-124 controls human vascular smooth muscle cell phenotypic switch via Sp1. Am J Physiol Heart Circ Physiol. 313:H641–H649. 2017. View Article : Google Scholar : PubMed/NCBI
18	Ahmadi A, Khansarinejad B, Hosseinkhani S, Ghanei M and Mowla SJ: miR-199a-5p and miR-495 target GRP78 within UPR pathway of lung cancer. Gene. 620:15–22. 2017. View Article : Google Scholar : PubMed/NCBI
19	Ma S, Jia W and Ni S: miR-199a-5p inhibits the progression of papillary thyroid carcinoma by targeting SNAI1. Biochem Biophys Res Commun. 497:181–186. 2018. View Article : Google Scholar : PubMed/NCBI
20	Zhu QD, Zhou QQ, Dong L, Huang Z, Wu F and Deng X: MiR-199a-5p inhibits the growth and metastasis of colorectal cancer cells by targeting ROCK1. Technol Cancer Res Treat. 17:15330346187755092018. View Article : Google Scholar : PubMed/NCBI
21	Qu D, Yang Y and Huang X: miR-199a-5p promotes proliferation and metastasis and epithelial-mesenchymal transition through targeting PIAS3 in cervical carcinoma. J Cell Biochem. 120:13562–13572. 2019. View Article : Google Scholar : PubMed/NCBI
22	Lim CS and Davies AH: Pathogenesis of primary varicose veins. Br J Surg. 96:1231–1242. 2009. View Article : Google Scholar : PubMed/NCBI
23	Pham TND, Perez White BE, Zhao H, Mortazavi F and Tonetti DA: Protein kinase C α enhances migration of breast cancer cells through FOXC2-mediated repression of p120-catenin. BMC Cancer. 17:8322017. View Article : Google Scholar : PubMed/NCBI
24	Ren YH, Liu KJ, Wang M, Yu YN, Yang K, Chen Q, Yu B, Wang W, Li QW, Wang J, et al: De-SUMOylation of FOXC2 by SENP3 promotes the epithelial-mesenchymal transition in gastric cancer cells. Oncotarget. 5:7093–7104. 2014. View Article : Google Scholar : PubMed/NCBI
25	Zheng CH, Quan Y, Li YY, Deng WG, Shao WJ and Fu Y: Expression of transcription factor FOXC2 in cervical cancer and effects of silencing on cervical cancer cell proliferation. Asian Pac J Cancer Prev. 15:1589–1595. 2014. View Article : Google Scholar : PubMed/NCBI
26	Liu B, Han SM, Tang XY, Han L and Li CZ: Overexpressed FOXC2 in ovarian cancer enhances the epithelial-to-mesenchymal transition and invasion of ovarian cancer cells. Oncol Rep. 31:2545–2554. 2014. View Article : Google Scholar : PubMed/NCBI
27	Wu X and Liu NF: FOXC2 transcription factor: A novel regulator of lymphangiogenesis. Lymphology. 44:35–41. 2011.PubMed/NCBI
28	Sano H, Leboeuf JP, Novitskiy SV, Seo S, Zaja-Milatovic S, Dikov MM and Kume T: The Foxc2 transcription factor regulates tumor angiogenesis. Biochem Biophys Res Commun. 392:201–206. 2010. View Article : Google Scholar : PubMed/NCBI
29	Zhang C, Li H and Guo X: FOXC2-AS1 regulates phenotypic transition, proliferation and migration of human great saphenous vein smooth muscle cells. Biol Res. 52:592019. View Article : Google Scholar : PubMed/NCBI