MiR-214 Regulates the Human Hair Follicle Stem Cell Proliferation and Differentiation by Targeting EZH2 and Wnt/β-Catenin Signaling Way In Vitro

Original Article
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Abstract

miR-214 plays a major role in the self-renewal of skin tissue. However, whether miR-214 regulates the proliferation and differentiation of human hair follicle stem cells (HFSCs) is unknown. Primary HFSCs were isolated from human scalp skin tissue, cultured, and identified using flow cytometry. An miR-214 mimic and inhibitor were constructed for transfection into HFSCs. The MTS and colony formation assays examined cell proliferation. Immunofluorescence detected the localization and expression levels of TCF4, β-catenin, and differentiation markers. Luciferase reporter and TOP/FOP Flash assays investigated whether miR-214 targeted EZH2 and regulated the Wnt/β-catenin signaling pathway. Western blot determined the expression levels of enhancer of zeste homolog 2 (EZH2), Wnt/β-catenin signaling-related proteins, and HFSC differentiation markers in cells subjected to miR-214 transfection. miR-214 expression was remarkably decreased during the proliferation and differentiation of HFSCs into transit-amplifying (TA) cells. Downregulation of miR-214 promotes the proliferation and differentiation of HFSCs. Overexpression of miR-214 led to decreased expression of EZH2, β-catenin, and TCF-4, whereas downregulation of miR-214 resulted in increased expression of EZH2, β-catenin, and TCF-4 as well as TA differentiation markers. Immunofluorescence assay revealed that inhibiting miR-214 triggered the entry of β-catenin and TCF-4 into the nucleus. The luciferase reporter and TOP/FOP Flash assays demonstrated that miR-214 directly targets EZH2 and affects Wnt/β-catenin signaling. The miR-214/EZH2/β-catenin axis could be considered a candidate target in tissue engineering and regenerative medicine for HFSCs.

Keywords

miR-214 Hair follicles stem cells Transit-amplifying cells EZH2 Wnt/β-catenin signal 

Notes

Acknowledgement

Funding was provided by the National Natural Science Foundation of China (81372114).

Compliance with ethical standards

Conflicts of interest

The authors have no financial conflicts of interest.

Ethical approval

The study was approved by the Ethical Committee of ChenZhou No.1 People’s Hospital (CZSDYRMYY-2017-18). All the donators signed the informed consent. There are no animal experiments carried out for this article.

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Copyright information

© The Korean Tissue Engineering and Regenerative Medicine Society and Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Rehabilitation, The First Affiliated HospitalJinan UniversityGuangzhouChina
  2. 2.Department of Rehabilitation, Chenzhou NO.1 People’s HospitalChenzhouChina
  3. 3.Department of Plastic Surgery, The 3rd Xiangya HospitalCentral South UniversityChangshaChina
  4. 4.Department of Rehabilitation Medicine, Guangdong Geriatric InstituteGuangdong General Hospital & Guangdong Academy of Medical SciencesGuangzhouChina

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