Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.

<h4>Background</h4>Cancer Stem Cells (CSCs) hypothesis asserts that only a small subset of cells within a tumour is capable of both tumour initiation and sustainment. The Epithelial-Mesenchymal Transition (EMT) is an embryonic developmental program that is often activated during cancer i...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Giuseppe Pirozzi, Virginia Tirino, Rosa Camerlingo, Renato Franco, Aantonello La Rocca, Eleonora Liguori, Nicola Martucci, Francesca Paino, Nicola Normanno, Gaetano Rocco
Formato: article
Lenguaje:EN
Publicado: Public Library of Science (PLoS) 2011
Materias:
R
Q
Acceso en línea:https://doaj.org/article/ac36782bc1994a128eba88bafa3e9aae
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
id oai:doaj.org-article:ac36782bc1994a128eba88bafa3e9aae
record_format dspace
spelling oai:doaj.org-article:ac36782bc1994a128eba88bafa3e9aae2021-11-18T06:50:54ZEpithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.1932-620310.1371/journal.pone.0021548https://doaj.org/article/ac36782bc1994a128eba88bafa3e9aae2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21738704/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Cancer Stem Cells (CSCs) hypothesis asserts that only a small subset of cells within a tumour is capable of both tumour initiation and sustainment. The Epithelial-Mesenchymal Transition (EMT) is an embryonic developmental program that is often activated during cancer invasion and metastasis. The aim of this study is to shed light on the relationship between EMT and CSCs by using LC31 lung cancer primary cell line.<h4>Materials and methods</h4>A549 and LC31 cell lines were treated with 2 ng/ml TGFβ-1 for 30 days, and 80 days, respectively. To evaluate EMT, morphological changes were assessed by light microscopy, immunofluorescence and cytometry for following markers: cytokeratins, e-cadherin, CD326 (epithelial markers) and CD90, and vimentin (mesenchymal markers). Moreover, RT-PCR for Slug, Twist and β-catenin genes were performed. On TGFβ-1 treated and untreated LC31 cell lines, we performed stemness tests such as pneumospheres growth and stem markers expression such as Oct4, Nanog, Sox2, c-kit and CD133. Western Blot for CD133 and tumorigenicity assays using NOD/SCID mice were performed.<h4>Results</h4>TGFβ-1 treated LC31 cell line lost its epithelial morphology assuming a fibroblast-like appearance. The same results were obtained for the A549 cell line (as control). Immunofluorescence and cytometry showed up-regulation of vimentin and CD90 and down-regulation of cytocheratin, e-cadherin and CD326 in TGFβ-1 treated LC31 and A549 cell lines. Slug, Twist and β-catenin m-RNA transcripts were up-regulated in TGFβ-1 treated LC31 cell line confirming EMT. This cell line showed also over-expression of Oct4, Nanog, Sox2 and CD133, all genes of stemness. In addition, in TGFβ-1 treated LC31 cell line, an increased pneumosphere-forming capacity and tumours-forming ability in NOD/SCID mice were detectable.<h4>Conclusions</h4>The induction of EMT by TGFβ-1 exposure, in primary lung cancer cell line results in the acquisition of mesenchymal profile and in the expression of stem cell markers.Giuseppe PirozziVirginia TirinoRosa CamerlingoRenato FrancoAantonello La RoccaEleonora LiguoriNicola MartucciFrancesca PainoNicola NormannoGaetano RoccoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 6, p e21548 (2011)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Giuseppe Pirozzi
Virginia Tirino
Rosa Camerlingo
Renato Franco
Aantonello La Rocca
Eleonora Liguori
Nicola Martucci
Francesca Paino
Nicola Normanno
Gaetano Rocco
Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
description <h4>Background</h4>Cancer Stem Cells (CSCs) hypothesis asserts that only a small subset of cells within a tumour is capable of both tumour initiation and sustainment. The Epithelial-Mesenchymal Transition (EMT) is an embryonic developmental program that is often activated during cancer invasion and metastasis. The aim of this study is to shed light on the relationship between EMT and CSCs by using LC31 lung cancer primary cell line.<h4>Materials and methods</h4>A549 and LC31 cell lines were treated with 2 ng/ml TGFβ-1 for 30 days, and 80 days, respectively. To evaluate EMT, morphological changes were assessed by light microscopy, immunofluorescence and cytometry for following markers: cytokeratins, e-cadherin, CD326 (epithelial markers) and CD90, and vimentin (mesenchymal markers). Moreover, RT-PCR for Slug, Twist and β-catenin genes were performed. On TGFβ-1 treated and untreated LC31 cell lines, we performed stemness tests such as pneumospheres growth and stem markers expression such as Oct4, Nanog, Sox2, c-kit and CD133. Western Blot for CD133 and tumorigenicity assays using NOD/SCID mice were performed.<h4>Results</h4>TGFβ-1 treated LC31 cell line lost its epithelial morphology assuming a fibroblast-like appearance. The same results were obtained for the A549 cell line (as control). Immunofluorescence and cytometry showed up-regulation of vimentin and CD90 and down-regulation of cytocheratin, e-cadherin and CD326 in TGFβ-1 treated LC31 and A549 cell lines. Slug, Twist and β-catenin m-RNA transcripts were up-regulated in TGFβ-1 treated LC31 cell line confirming EMT. This cell line showed also over-expression of Oct4, Nanog, Sox2 and CD133, all genes of stemness. In addition, in TGFβ-1 treated LC31 cell line, an increased pneumosphere-forming capacity and tumours-forming ability in NOD/SCID mice were detectable.<h4>Conclusions</h4>The induction of EMT by TGFβ-1 exposure, in primary lung cancer cell line results in the acquisition of mesenchymal profile and in the expression of stem cell markers.
format article
author Giuseppe Pirozzi
Virginia Tirino
Rosa Camerlingo
Renato Franco
Aantonello La Rocca
Eleonora Liguori
Nicola Martucci
Francesca Paino
Nicola Normanno
Gaetano Rocco
author_facet Giuseppe Pirozzi
Virginia Tirino
Rosa Camerlingo
Renato Franco
Aantonello La Rocca
Eleonora Liguori
Nicola Martucci
Francesca Paino
Nicola Normanno
Gaetano Rocco
author_sort Giuseppe Pirozzi
title Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
title_short Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
title_full Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
title_fullStr Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
title_full_unstemmed Epithelial to mesenchymal transition by TGFβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
title_sort epithelial to mesenchymal transition by tgfβ-1 induction increases stemness characteristics in primary non small cell lung cancer cell line.
publisher Public Library of Science (PLoS)
publishDate 2011
url https://doaj.org/article/ac36782bc1994a128eba88bafa3e9aae
work_keys_str_mv AT giuseppepirozzi epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT virginiatirino epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT rosacamerlingo epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT renatofranco epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT aantonellolarocca epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT eleonoraliguori epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT nicolamartucci epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT francescapaino epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT nicolanormanno epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
AT gaetanorocco epithelialtomesenchymaltransitionbytgfb1inductionincreasesstemnesscharacteristicsinprimarynonsmallcelllungcancercellline
_version_ 1718424313238913024