Cargando…

Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features

Thymosin β4 (Tβ4) is a small, 44-amino acid polypeptide. It has been implicated in multiple processes, including cell movement, angiogenesis, and stemness. Previously, we reported that melanoma cell lines differ in Tβ4 levels. Studies on stable clones with silenced TMSB4X expression showed that Tβ4...

Descripción completa

Detalles Bibliográficos
Autores principales: Makowiecka, Aleksandra, Mazurkiewicz, Ewa, Mrówczyńska, Ewa, Malek, Natalia, Battistella, Alice, Lazzarino, Marco, Nowak, Dorota, Mazur, Antonina Joanna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8067020/
https://www.ncbi.nlm.nih.gov/pubmed/33807338
http://dx.doi.org/10.3390/cells10040769
_version_ 1783682704073555968
author Makowiecka, Aleksandra
Mazurkiewicz, Ewa
Mrówczyńska, Ewa
Malek, Natalia
Battistella, Alice
Lazzarino, Marco
Nowak, Dorota
Mazur, Antonina Joanna
author_facet Makowiecka, Aleksandra
Mazurkiewicz, Ewa
Mrówczyńska, Ewa
Malek, Natalia
Battistella, Alice
Lazzarino, Marco
Nowak, Dorota
Mazur, Antonina Joanna
author_sort Makowiecka, Aleksandra
collection PubMed
description Thymosin β4 (Tβ4) is a small, 44-amino acid polypeptide. It has been implicated in multiple processes, including cell movement, angiogenesis, and stemness. Previously, we reported that melanoma cell lines differ in Tβ4 levels. Studies on stable clones with silenced TMSB4X expression showed that Tβ4 impacted adhesion and epithelial-mesenchymal transition progression. Here, we show that the cells with silenced TMSB4X expression exhibited altered actin cytoskeleton’s organization and subcellular relocalization of two intermediate filament proteins: Nestin and Vimentin. The rearrangement of the cell cytoskeleton resulted in changes in the cells’ topology, height, and stiffness defined by Young’s modulus. Simultaneously, only for some A375 clones with a lowered Tβ4 level, we observed a decreased ability to initiate colony formation in soft agar, tumor formation in vivo, and alterations in Nanog’s expression level transcription factor regulating stemness. Thus, we show for the first time that in A375 cells, biomechanical properties are not directly coupled to stemness features, and this cell line is phenotypically heterogeneous.
format Online
Article
Text
id pubmed-8067020
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-80670202021-04-25 Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features Makowiecka, Aleksandra Mazurkiewicz, Ewa Mrówczyńska, Ewa Malek, Natalia Battistella, Alice Lazzarino, Marco Nowak, Dorota Mazur, Antonina Joanna Cells Article Thymosin β4 (Tβ4) is a small, 44-amino acid polypeptide. It has been implicated in multiple processes, including cell movement, angiogenesis, and stemness. Previously, we reported that melanoma cell lines differ in Tβ4 levels. Studies on stable clones with silenced TMSB4X expression showed that Tβ4 impacted adhesion and epithelial-mesenchymal transition progression. Here, we show that the cells with silenced TMSB4X expression exhibited altered actin cytoskeleton’s organization and subcellular relocalization of two intermediate filament proteins: Nestin and Vimentin. The rearrangement of the cell cytoskeleton resulted in changes in the cells’ topology, height, and stiffness defined by Young’s modulus. Simultaneously, only for some A375 clones with a lowered Tβ4 level, we observed a decreased ability to initiate colony formation in soft agar, tumor formation in vivo, and alterations in Nanog’s expression level transcription factor regulating stemness. Thus, we show for the first time that in A375 cells, biomechanical properties are not directly coupled to stemness features, and this cell line is phenotypically heterogeneous. MDPI 2021-03-31 /pmc/articles/PMC8067020/ /pubmed/33807338 http://dx.doi.org/10.3390/cells10040769 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
Makowiecka, Aleksandra
Mazurkiewicz, Ewa
Mrówczyńska, Ewa
Malek, Natalia
Battistella, Alice
Lazzarino, Marco
Nowak, Dorota
Mazur, Antonina Joanna
Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features
title Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features
title_full Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features
title_fullStr Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features
title_full_unstemmed Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features
title_short Changes in Biomechanical Properties of A375 Cells Due to the Silencing of TMSB4X Expression Are Not Directly Correlated with Alterations in Their Stemness Features
title_sort changes in biomechanical properties of a375 cells due to the silencing of tmsb4x expression are not directly correlated with alterations in their stemness features
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8067020/
https://www.ncbi.nlm.nih.gov/pubmed/33807338
http://dx.doi.org/10.3390/cells10040769
work_keys_str_mv AT makowieckaaleksandra changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures
AT mazurkiewiczewa changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures
AT mrowczynskaewa changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures
AT maleknatalia changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures
AT battistellaalice changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures
AT lazzarinomarco changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures
AT nowakdorota changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures
AT mazurantoninajoanna changesinbiomechanicalpropertiesofa375cellsduetothesilencingoftmsb4xexpressionarenotdirectlycorrelatedwithalterationsintheirstemnessfeatures