Cargando…

Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases

In passages above ten and growing very actively, we observed that some human lung fibroblasts cultured under standard conditions were transformed into a lineage of epithelial-like cells (ELC). To systematically evaluate the possible mesenchymal–epithelial transition (MET) occurrence, fibroblasts wer...

Descripción completa

Detalles Bibliográficos
Autores principales: Becerril, Carina, Montaño, Martha, Cisneros, José, Mendoza-Milla, Criselda, Pardo, Annie, Ortiz-Quintero, Blanca, Selman, Moisés, Ramos, Carlos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8000192/
https://www.ncbi.nlm.nih.gov/pubmed/33806618
http://dx.doi.org/10.3390/biom11030378
_version_ 1783670947990994944
author Becerril, Carina
Montaño, Martha
Cisneros, José
Mendoza-Milla, Criselda
Pardo, Annie
Ortiz-Quintero, Blanca
Selman, Moisés
Ramos, Carlos
author_facet Becerril, Carina
Montaño, Martha
Cisneros, José
Mendoza-Milla, Criselda
Pardo, Annie
Ortiz-Quintero, Blanca
Selman, Moisés
Ramos, Carlos
author_sort Becerril, Carina
collection PubMed
description In passages above ten and growing very actively, we observed that some human lung fibroblasts cultured under standard conditions were transformed into a lineage of epithelial-like cells (ELC). To systematically evaluate the possible mesenchymal–epithelial transition (MET) occurrence, fibroblasts were obtained from normal lungs and also from lungs affected by idiopathic interstitial diseases. When an unusual epithelial-like phenotypic change was observed, cultured cells were characterized by confocal immunofluorescence microscopy, immunoblotting, immunocytochemistry, cytofluorometry, gelatin zymography, RT-qPCR, and hybridization in a whole-transcript human microarray. Additionally, microvesicles fraction (MVs) from ELC and fibroblasts were used to induce MET, while the microRNAs (miRNAs) contained in the MVs were identified. Pattern-gene expression of the original fibroblasts and the derived ELC revealed profound changes, upregulating characteristic epithelial-cell genes and downregulating mesenchymal genes, with a marked increase of E-cadherin, cytokeratin, and ZO-1, and the loss of expression of α-SMA, collagen type I, and Thy-1 cell surface antigen (CD90). Fibroblasts, exposed to culture media or MVs from the ELC, acquired ELC phenotype. The miRNAs in MVs shown six expressed exclusively in fibroblasts, and three only in ELC; moreover, twelve miRNAs were differentially expressed between fibroblasts and ELC, all of them but one was overexpressed in fibroblasts. These findings suggest that the MET-like process can occur in human lung fibroblasts, either from normal or diseased lungs. However, the biological implication is unclear.
format Online
Article
Text
id pubmed-8000192
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-80001922021-03-28 Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases Becerril, Carina Montaño, Martha Cisneros, José Mendoza-Milla, Criselda Pardo, Annie Ortiz-Quintero, Blanca Selman, Moisés Ramos, Carlos Biomolecules Article In passages above ten and growing very actively, we observed that some human lung fibroblasts cultured under standard conditions were transformed into a lineage of epithelial-like cells (ELC). To systematically evaluate the possible mesenchymal–epithelial transition (MET) occurrence, fibroblasts were obtained from normal lungs and also from lungs affected by idiopathic interstitial diseases. When an unusual epithelial-like phenotypic change was observed, cultured cells were characterized by confocal immunofluorescence microscopy, immunoblotting, immunocytochemistry, cytofluorometry, gelatin zymography, RT-qPCR, and hybridization in a whole-transcript human microarray. Additionally, microvesicles fraction (MVs) from ELC and fibroblasts were used to induce MET, while the microRNAs (miRNAs) contained in the MVs were identified. Pattern-gene expression of the original fibroblasts and the derived ELC revealed profound changes, upregulating characteristic epithelial-cell genes and downregulating mesenchymal genes, with a marked increase of E-cadherin, cytokeratin, and ZO-1, and the loss of expression of α-SMA, collagen type I, and Thy-1 cell surface antigen (CD90). Fibroblasts, exposed to culture media or MVs from the ELC, acquired ELC phenotype. The miRNAs in MVs shown six expressed exclusively in fibroblasts, and three only in ELC; moreover, twelve miRNAs were differentially expressed between fibroblasts and ELC, all of them but one was overexpressed in fibroblasts. These findings suggest that the MET-like process can occur in human lung fibroblasts, either from normal or diseased lungs. However, the biological implication is unclear. MDPI 2021-03-04 /pmc/articles/PMC8000192/ /pubmed/33806618 http://dx.doi.org/10.3390/biom11030378 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 (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Becerril, Carina
Montaño, Martha
Cisneros, José
Mendoza-Milla, Criselda
Pardo, Annie
Ortiz-Quintero, Blanca
Selman, Moisés
Ramos, Carlos
Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases
title Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases
title_full Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases
title_fullStr Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases
title_full_unstemmed Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases
title_short Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases
title_sort mesenchymal–epithelial transition in fibroblasts of human normal lungs and interstitial lung diseases
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8000192/
https://www.ncbi.nlm.nih.gov/pubmed/33806618
http://dx.doi.org/10.3390/biom11030378
work_keys_str_mv AT becerrilcarina mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases
AT montanomartha mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases
AT cisnerosjose mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases
AT mendozamillacriselda mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases
AT pardoannie mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases
AT ortizquinteroblanca mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases
AT selmanmoises mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases
AT ramoscarlos mesenchymalepithelialtransitioninfibroblastsofhumannormallungsandinterstitiallungdiseases