Cargando…
Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells
Transparency of the human cornea is responsible for clear vision, which is maintained by a monolayer of non-proliferative human corneal endothelial cells (HCEnCs). Dysfunction of these cells can result in irreversible corneal blindness. It is important to identify key factors that limit the prolifer...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10380751/ https://www.ncbi.nlm.nih.gov/pubmed/37511249 http://dx.doi.org/10.3390/ijms241411490 |
_version_ | 1785080273220665344 |
---|---|
author | Parekh, Mohit Ramos, Tiago Ferrari, Stefano Ahmad, Sajjad |
author_facet | Parekh, Mohit Ramos, Tiago Ferrari, Stefano Ahmad, Sajjad |
author_sort | Parekh, Mohit |
collection | PubMed |
description | Transparency of the human cornea is responsible for clear vision, which is maintained by a monolayer of non-proliferative human corneal endothelial cells (HCEnCs). Dysfunction of these cells can result in irreversible corneal blindness. It is important to identify key factors that limit the proliferation of HCEnCs and thus attempt to reverse them. Extracellular vesicles contain cargo which includes microRNAs (miRNAs) that can modulate a cellular function. In non small cell lung cancer, expression of miR-195-5p has been shown to inhibit proliferation; therefore, we aimed to investigate the inhibitory effect of miR-195-5p in inducing the proliferation of HCEnCs. Human corneal endothelial cell line (HCEC-12) and primary HCEnCs were cultured with miR-195-5p scramble, mimic or inhibitor. Corneal tissues from human cadaveric and FECD donors, and from pigs, mice and rabbits, were used for RT-PCR. miR-195-5p showed an abundance value of 11,363.31 a.u. When normalized against HCEnCs from cadaveric donors, FECD tissues showed a significant upregulation of miR-195-5p (p < 0.05) but was significantly downregulated in pig (p < 0.001), mouse (p < 0.01) and rabbit (p < 0.001) CEnCs, which have known proliferative capacity. Proliferation, cell doubling, and wound healing rates were significantly higher when miR-195-5p was inhibited. Inhibiting miR-195-5p showed a significant improvement in viability (HEC staining), decreased cell apoptosis (TdT-dNTP staining) and expression of ZO-1, NA(+)/K(+)-ATPase and Ki-67 markers. Expression of miR-195-5p is found in HCEnCs and FECD cells, which restricts the proliferation of these cells. However, inhibiting miR-195-5p can induce the proliferation of HCEnCs, which opens exciting directions for future research in prolonging FECD pathogenesis by increasing the proliferative capacity of HCEnCs using anti-miR therapy in vivo. |
format | Online Article Text |
id | pubmed-10380751 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-103807512023-07-29 Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells Parekh, Mohit Ramos, Tiago Ferrari, Stefano Ahmad, Sajjad Int J Mol Sci Article Transparency of the human cornea is responsible for clear vision, which is maintained by a monolayer of non-proliferative human corneal endothelial cells (HCEnCs). Dysfunction of these cells can result in irreversible corneal blindness. It is important to identify key factors that limit the proliferation of HCEnCs and thus attempt to reverse them. Extracellular vesicles contain cargo which includes microRNAs (miRNAs) that can modulate a cellular function. In non small cell lung cancer, expression of miR-195-5p has been shown to inhibit proliferation; therefore, we aimed to investigate the inhibitory effect of miR-195-5p in inducing the proliferation of HCEnCs. Human corneal endothelial cell line (HCEC-12) and primary HCEnCs were cultured with miR-195-5p scramble, mimic or inhibitor. Corneal tissues from human cadaveric and FECD donors, and from pigs, mice and rabbits, were used for RT-PCR. miR-195-5p showed an abundance value of 11,363.31 a.u. When normalized against HCEnCs from cadaveric donors, FECD tissues showed a significant upregulation of miR-195-5p (p < 0.05) but was significantly downregulated in pig (p < 0.001), mouse (p < 0.01) and rabbit (p < 0.001) CEnCs, which have known proliferative capacity. Proliferation, cell doubling, and wound healing rates were significantly higher when miR-195-5p was inhibited. Inhibiting miR-195-5p showed a significant improvement in viability (HEC staining), decreased cell apoptosis (TdT-dNTP staining) and expression of ZO-1, NA(+)/K(+)-ATPase and Ki-67 markers. Expression of miR-195-5p is found in HCEnCs and FECD cells, which restricts the proliferation of these cells. However, inhibiting miR-195-5p can induce the proliferation of HCEnCs, which opens exciting directions for future research in prolonging FECD pathogenesis by increasing the proliferative capacity of HCEnCs using anti-miR therapy in vivo. MDPI 2023-07-15 /pmc/articles/PMC10380751/ /pubmed/37511249 http://dx.doi.org/10.3390/ijms241411490 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Parekh, Mohit Ramos, Tiago Ferrari, Stefano Ahmad, Sajjad Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells |
title | Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells |
title_full | Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells |
title_fullStr | Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells |
title_full_unstemmed | Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells |
title_short | Inhibiting miR-195-5p Induces Proliferation of Human Corneal Endothelial Cells |
title_sort | inhibiting mir-195-5p induces proliferation of human corneal endothelial cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10380751/ https://www.ncbi.nlm.nih.gov/pubmed/37511249 http://dx.doi.org/10.3390/ijms241411490 |
work_keys_str_mv | AT parekhmohit inhibitingmir1955pinducesproliferationofhumancornealendothelialcells AT ramostiago inhibitingmir1955pinducesproliferationofhumancornealendothelialcells AT ferraristefano inhibitingmir1955pinducesproliferationofhumancornealendothelialcells AT ahmadsajjad inhibitingmir1955pinducesproliferationofhumancornealendothelialcells |