Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B

Rosso, Marina; Lapyckyj, Lara; Besso, María José; Monge i Azemar, Marta; Reventós Puigjaner, Jaume; Canals, Francesc; Quevedo Cuenca, Jorge Oswaldo; Matos, María Laura; Hebe Vazquez Levin, Mónica

Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B

Breast cancer (BC) is the most common female cancer and the leading cause of cancer death in women worldwide. Alterations in epithelial cadherin (E-cadherin) expression and functions are associated to BC, but the underlying molecular mechanisms have not been fully elucidated. We have previously repo...

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Autores:	Rosso, Marina, Lapyckyj, Lara, Besso, María José, Monge i Azemar, Marta, Reventós Puigjaner, Jaume, Canals, Francesc, Quevedo Cuenca, Jorge Oswaldo, Matos, María Laura, Hebe Vazquez Levin, Mónica
Formato:	artículo
Estado:	Versión publicada
Fecha de publicación:	2019
País:	España
Recursos:	Universidad de Barcelona
Repositorio:	Dipòsit Digital de la UB
OAI Identifier:	oai:diposit.ub.edu:2445/177925
Acesso em linha:	https://hdl.handle.net/2445/177925
Access Level:	acceso abierto
Palavra-chave:	Càncer de mama Tumors Patologia cel·lular Breast cancer Cellular pathology

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spelling	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase BRosso, MarinaLapyckyj, LaraBesso, María JoséMonge i Azemar, MartaReventós Puigjaner, JaumeCanals, FrancescQuevedo Cuenca, Jorge OswaldoMatos, María LauraHebe Vazquez Levin, MónicaCàncer de mamaTumorsPatologia cel·lularBreast cancerTumorsCellular pathologyBreast cancer (BC) is the most common female cancer and the leading cause of cancer death in women worldwide. Alterations in epithelial cadherin (E-cadherin) expression and functions are associated to BC, but the underlying molecular mechanisms have not been fully elucidated. We have previously reported a novel human E-cadherin splice variant (E-cadherin variant) mRNA. Stable transfectants in MCF-7 human BC cells (MCF7Ecadvar) depicted fibroblast-like cell morphology, E-cadherin wild-type downregulation, and other molecular changes characteristic of the epithelial-to-mesenchymal transition process, reduced cell-cell adhesion, and increased cell migration and invasion. In this study, a two-dimensional differential gel electrophoresis (2D-DIGE) combined with mass spectrometry (MS) protein identification and bioinformatics analyses were Results: By 2D-DIGE and MS analysis, 50 proteins were found differentially expressed (≥ Δ1.5) in MCF7Ecadvar compared to control cells. Validation of transcript expression was done in the ten most overexpressed and underexpressed proteins. Bioinformatics analyses revealed that 39 of the 50 proteins identified had been previously associated to BC. Moreover, metabolic processes were the most affected, and glycolysis the canonical pathway most altered. The lactate dehydrogenase B (LDHB) was the highest overexpressed protein, and transcript levels were higher in MCF7Ecadvar than in control cells. In agreement with these findings, MCF7Ecadvar conditioned media had lower glucose and higher lactate levels than control cells. MCF7Ecadvar cell treatment with 5 mM of the glycolytic inhibitor 2-deoxy-glucose led to decreased cell viability, and modulation of LDHB expression in MCF7Ecadvar cells with a specific small interfering RNA resulted in decreased cell proliferation. Finally, a positive association between expression levels of the E-cadherin variant and LDHB transcripts was demonstrated in 21 human breast tumor tissues, and breast tumor samples with higher Ki67 expression showed higher LDHB mRNA levels. Conclusions: Results from this investigation contributed to further characterize molecular changes associated to the novel E-cadherin splice variant expression in BC cells. They also revealed an association between expression of the novel variant and changes related to BC progression and aggressiveness, in particular those associated to cell metabolism. Keywords: Breast cancer, Epithelial cadherin, Alternative splicing, Epithelial to mesenchymal transition, Proteomic analysis, 2D-DIGE, Mass spectrometry, Glycolysis, Lactate dehydrogenase BSpringer Nature2019info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/177925Articles publicats en revistes (Farmàcia, Tecnologia Farmacèutica i Fisicoquímica)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.1186/s40170-019-0196-9Cancer & Metabolism, 2019, vol. 7, num. 5https://doi.org/10.1186/s40170-019-0196-9info:eu-repo/grantAgreement/EC/FP7/269285cc-by (c) Rosso, Marina et al., 2019https://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/1779252026-05-27T06:46:51Z
dc.title.none.fl_str_mv	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B
title	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B
spellingShingle	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B Rosso, Marina Càncer de mama Tumors Patologia cel·lular Breast cancer Tumors Cellular pathology
title_short	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B
title_full	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B
title_fullStr	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B
title_full_unstemmed	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B
title_sort	Characterization of the molecular changes associated with the overexpression of a novel epithelial cadherin splice variant mRNA in a breast cancer model using proteomics and bioinformatics approaches: identification of changes in cell metabolism and an increased expression of lactate dehydrogenase B
dc.creator.none.fl_str_mv	Rosso, Marina Lapyckyj, Lara Besso, María José Monge i Azemar, Marta Reventós Puigjaner, Jaume Canals, Francesc Quevedo Cuenca, Jorge Oswaldo Matos, María Laura Hebe Vazquez Levin, Mónica
author	Rosso, Marina
author_facet	Rosso, Marina Lapyckyj, Lara Besso, María José Monge i Azemar, Marta Reventós Puigjaner, Jaume Canals, Francesc Quevedo Cuenca, Jorge Oswaldo Matos, María Laura Hebe Vazquez Levin, Mónica
author_role	author
author2	Lapyckyj, Lara Besso, María José Monge i Azemar, Marta Reventós Puigjaner, Jaume Canals, Francesc Quevedo Cuenca, Jorge Oswaldo Matos, María Laura Hebe Vazquez Levin, Mónica
author2_role	author author author author author author author author
dc.subject.none.fl_str_mv	Càncer de mama Tumors Patologia cel·lular Breast cancer Tumors Cellular pathology
topic	Càncer de mama Tumors Patologia cel·lular Breast cancer Tumors Cellular pathology
description	Breast cancer (BC) is the most common female cancer and the leading cause of cancer death in women worldwide. Alterations in epithelial cadherin (E-cadherin) expression and functions are associated to BC, but the underlying molecular mechanisms have not been fully elucidated. We have previously reported a novel human E-cadherin splice variant (E-cadherin variant) mRNA. Stable transfectants in MCF-7 human BC cells (MCF7Ecadvar) depicted fibroblast-like cell morphology, E-cadherin wild-type downregulation, and other molecular changes characteristic of the epithelial-to-mesenchymal transition process, reduced cell-cell adhesion, and increased cell migration and invasion. In this study, a two-dimensional differential gel electrophoresis (2D-DIGE) combined with mass spectrometry (MS) protein identification and bioinformatics analyses were Results: By 2D-DIGE and MS analysis, 50 proteins were found differentially expressed (≥ Δ1.5) in MCF7Ecadvar compared to control cells. Validation of transcript expression was done in the ten most overexpressed and underexpressed proteins. Bioinformatics analyses revealed that 39 of the 50 proteins identified had been previously associated to BC. Moreover, metabolic processes were the most affected, and glycolysis the canonical pathway most altered. The lactate dehydrogenase B (LDHB) was the highest overexpressed protein, and transcript levels were higher in MCF7Ecadvar than in control cells. In agreement with these findings, MCF7Ecadvar conditioned media had lower glucose and higher lactate levels than control cells. MCF7Ecadvar cell treatment with 5 mM of the glycolytic inhibitor 2-deoxy-glucose led to decreased cell viability, and modulation of LDHB expression in MCF7Ecadvar cells with a specific small interfering RNA resulted in decreased cell proliferation. Finally, a positive association between expression levels of the E-cadherin variant and LDHB transcripts was demonstrated in 21 human breast tumor tissues, and breast tumor samples with higher Ki67 expression showed higher LDHB mRNA levels. Conclusions: Results from this investigation contributed to further characterize molecular changes associated to the novel E-cadherin splice variant expression in BC cells. They also revealed an association between expression of the novel variant and changes related to BC progression and aggressiveness, in particular those associated to cell metabolism. Keywords: Breast cancer, Epithelial cadherin, Alternative splicing, Epithelial to mesenchymal transition, Proteomic analysis, 2D-DIGE, Mass spectrometry, Glycolysis, Lactate dehydrogenase B
publishDate	2019
dc.date.none.fl_str_mv	2019
dc.type.none.fl_str_mv	info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion
format	article
status_str	publishedVersion
dc.identifier.none.fl_str_mv	https://hdl.handle.net/2445/177925
url	https://hdl.handle.net/2445/177925
dc.language.none.fl_str_mv	Inglés
language_invalid_str_mv	Inglés
dc.relation.none.fl_str_mv	Reproducció del document publicat a: https://doi.org/10.1186/s40170-019-0196-9 Cancer & Metabolism, 2019, vol. 7, num. 5 https://doi.org/10.1186/s40170-019-0196-9 info:eu-repo/grantAgreement/EC/FP7/269285
dc.rights.none.fl_str_mv	cc-by (c) Rosso, Marina et al., 2019 https://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess
rights_invalid_str_mv	cc-by (c) Rosso, Marina et al., 2019 https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv	openAccess
dc.format.none.fl_str_mv	application/pdf
dc.publisher.none.fl_str_mv	Springer Nature
publisher.none.fl_str_mv	Springer Nature
dc.source.none.fl_str_mv	Articles publicats en revistes (Farmàcia, Tecnologia Farmacèutica i Fisicoquímica) reponame:Dipòsit Digital de la UB instname:Universidad de Barcelona
instname_str	Universidad de Barcelona
reponame_str	Dipòsit Digital de la UB
collection	Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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