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miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1

Recent findings suggest that epithelial to mesenchymal transition (EMT), a key step during heart development, is involved in cardiac tissue repair following myocardial infarction (MI). MicroRNAs (miRNAs) act as key regulators in EMT processes; however, the mechanisms by which miRNAs target epicardia...

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Autores principales: Pontemezzo, Elena, Foglio, Eleonora, Vernucci, Enza, Magenta, Alessandra, D’Agostino, Marco, Sileno, Sara, Astanina, Elena, Bussolino, Federico, Pellegrini, Laura, Germani, Antonia, Russo, Matteo Antonio, Limana, Federica
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125323/
https://www.ncbi.nlm.nih.gov/pubmed/34067060
http://dx.doi.org/10.3390/ijms22094971
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author Pontemezzo, Elena
Foglio, Eleonora
Vernucci, Enza
Magenta, Alessandra
D’Agostino, Marco
Sileno, Sara
Astanina, Elena
Bussolino, Federico
Pellegrini, Laura
Germani, Antonia
Russo, Matteo Antonio
Limana, Federica
author_facet Pontemezzo, Elena
Foglio, Eleonora
Vernucci, Enza
Magenta, Alessandra
D’Agostino, Marco
Sileno, Sara
Astanina, Elena
Bussolino, Federico
Pellegrini, Laura
Germani, Antonia
Russo, Matteo Antonio
Limana, Federica
author_sort Pontemezzo, Elena
collection PubMed
description Recent findings suggest that epithelial to mesenchymal transition (EMT), a key step during heart development, is involved in cardiac tissue repair following myocardial infarction (MI). MicroRNAs (miRNAs) act as key regulators in EMT processes; however, the mechanisms by which miRNAs target epicardial EMT remain largely unknown. Here, by using an in vitro model of epicardial EMT, we investigated the role of miRNAs as regulators of this process and their potential targets. EMT was induced in murine epicardial-mesothelial cells (EMCs) through TGF β1 treatment for 48, 72, and 96 h as indicated by the expression of EMT-related genes by qRT-PCR, WB, and immunofluorescence. Further, enhanced expression of stemness genes was also detected. Among several EMT-related miRNAs, miR-200c-3p expression resulted as the most strongly suppressed. Interestingly, we also found a significant upregulation of Follistatin-related protein 1 (FSTL1), a miR-200c predicted target already identified as a potent cardiogenic factor produced by epicardial cells that promotes regeneration following MI. Dual-luciferase reporter assay demonstrated that miR-200c-3p directly targeted the 3′-untranslated region of FSTL1 in EMCs. Consistently, WB analysis showed that knockdown of miR-200c-3p significantly increased FSTL1 expression, whereas overexpression of miR-200c-3p counteracted TGF β1-mediated FSTL1 upregulation. Importantly, FSTL1 silencing maintained epithelial features in EMCs, despite EMT induction by TGF β1, and attenuated EMT-associated traits, including migration and stemness. In conclusion, epicardial FSTL1, an important cardiogenic factor in its secreted form, induces EMT, stemness, and migration of EMCs in a miR-200c-3p dependent pathway.
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spelling pubmed-81253232021-05-17 miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1 Pontemezzo, Elena Foglio, Eleonora Vernucci, Enza Magenta, Alessandra D’Agostino, Marco Sileno, Sara Astanina, Elena Bussolino, Federico Pellegrini, Laura Germani, Antonia Russo, Matteo Antonio Limana, Federica Int J Mol Sci Article Recent findings suggest that epithelial to mesenchymal transition (EMT), a key step during heart development, is involved in cardiac tissue repair following myocardial infarction (MI). MicroRNAs (miRNAs) act as key regulators in EMT processes; however, the mechanisms by which miRNAs target epicardial EMT remain largely unknown. Here, by using an in vitro model of epicardial EMT, we investigated the role of miRNAs as regulators of this process and their potential targets. EMT was induced in murine epicardial-mesothelial cells (EMCs) through TGF β1 treatment for 48, 72, and 96 h as indicated by the expression of EMT-related genes by qRT-PCR, WB, and immunofluorescence. Further, enhanced expression of stemness genes was also detected. Among several EMT-related miRNAs, miR-200c-3p expression resulted as the most strongly suppressed. Interestingly, we also found a significant upregulation of Follistatin-related protein 1 (FSTL1), a miR-200c predicted target already identified as a potent cardiogenic factor produced by epicardial cells that promotes regeneration following MI. Dual-luciferase reporter assay demonstrated that miR-200c-3p directly targeted the 3′-untranslated region of FSTL1 in EMCs. Consistently, WB analysis showed that knockdown of miR-200c-3p significantly increased FSTL1 expression, whereas overexpression of miR-200c-3p counteracted TGF β1-mediated FSTL1 upregulation. Importantly, FSTL1 silencing maintained epithelial features in EMCs, despite EMT induction by TGF β1, and attenuated EMT-associated traits, including migration and stemness. In conclusion, epicardial FSTL1, an important cardiogenic factor in its secreted form, induces EMT, stemness, and migration of EMCs in a miR-200c-3p dependent pathway. MDPI 2021-05-07 /pmc/articles/PMC8125323/ /pubmed/34067060 http://dx.doi.org/10.3390/ijms22094971 Text en © 2021 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
Pontemezzo, Elena
Foglio, Eleonora
Vernucci, Enza
Magenta, Alessandra
D’Agostino, Marco
Sileno, Sara
Astanina, Elena
Bussolino, Federico
Pellegrini, Laura
Germani, Antonia
Russo, Matteo Antonio
Limana, Federica
miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1
title miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1
title_full miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1
title_fullStr miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1
title_full_unstemmed miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1
title_short miR-200c-3p Regulates Epitelial-to-Mesenchymal Transition in Epicardial Mesothelial Cells by Targeting Epicardial Follistatin-Related Protein 1
title_sort mir-200c-3p regulates epitelial-to-mesenchymal transition in epicardial mesothelial cells by targeting epicardial follistatin-related protein 1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125323/
https://www.ncbi.nlm.nih.gov/pubmed/34067060
http://dx.doi.org/10.3390/ijms22094971
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