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Hsa-miR-150-5p inhibits Wnt-beta-catenin signaling in human corneal epithelial stem cells

Kalaimani, Lavanya; Devarajan, Bharanidharan; Namperumalsamy, Venkatesh Prajna; Veerappan, Muthukkaruppan; Daniels, Julie T; Chidambaranathan, Gowri Priya; (2022) Hsa-miR-150-5p inhibits Wnt-beta-catenin signaling in human corneal epithelial stem cells. Molecular Vision , 28 pp. 178-191. Green open access

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Abstract

Purpose: In our earlier study, we identified hsa-miR-150-5p as a highly expressed miRNA in enriched corneal epithelial stem cells (CESCs). In this study, we aimed to understand the molecular regulatory function of hsa-miR-150-5p in association with the maintenance of stemness in CESCs. Methods: The target mRNAs of hsa-miR-150-5p were predicted and subjected to pathway analysis to identify targets for functional studies. Primary cultured limbal epithelial cells were transfected with hsa-miR-150-5p mimic, inhibitor, or scrambled sequence using Lipofectamine 3000. The transfected cells were analyzed to determine (i) their colony-forming potential; (ii) the expression levels of stem cell (SC) markers/transcription factors (ABCG2, NANOG, OCT4, KLF4, and ΔNp63), the differentiation marker (Cx43), and the hsa-miR-150-5p predicted targets (JARID2, INHBA, AKT3, and CTNNB1) by qPCR; and (iii) the expression levels of ABCG2, p63α, Cx43, JARID2, AKT3, p-AKT3, β-catenin, and active β-catenin by immunofluorescence staining and/or western blotting. Results: The ectopic expression level of hsa-miR-150-5p increased the colony-forming potential (8.29% ± 0.47%, p < 0.001) with the ability to form holoclone-like colonies compared with the control (1.8% ± 0.47%). The mimic-treated cells had higher expression levels of the SC markers but reduced expression levels of Cx43 and the targets of hsa-miR-150-5p that are involved in the Wnt-β-catenin signaling pathway. The expression levels of β-catenin and active β-catenin in the inhibitor-transfected cells were higher than those in the control cells, and the localized nuclear expression indicated the activation of Wnt signaling. Conclusions: Our results indicate a regulatory role for hsa-miR-150-5p in the maintenance of CESCs by inhibiting the Wnt signaling pathway.

Type: Article
Title: Hsa-miR-150-5p inhibits Wnt-beta-catenin signaling in human corneal epithelial stem cells
Open access status: An open access version is available from UCL Discovery
Publisher version: http://www.molvis.org/molvis/v28/178/
Language: English
Additional information: For noncommercial use, republished material must include a citation to the original work in Molecular Vision and must not be changed from the original. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for license terms.
Keywords: Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Ophthalmology, EXPRESSING HIGH-LEVELS, SELF-RENEWAL, PATHWAY, SUBSET, TARGET, DIFFERENTIATION, MIR-150-5P, NICHE
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL
URI: https://discovery.ucl.ac.uk/id/eprint/10156469
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