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
Abstract Background 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...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-05-01
|
| Series: | Cancer & Metabolism |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40170-019-0196-9 |
| _version_ | 1828385578094166016 |
|---|---|
| author | Marina Rosso Lara Lapyckyj María José Besso Marta Monge Jaume Reventós Francesc Canals Jorge Oswaldo Quevedo Cuenca María Laura Matos Mónica Hebe Vazquez-Levin |
| author_facet | Marina Rosso Lara Lapyckyj María José Besso Marta Monge Jaume Reventós Francesc Canals Jorge Oswaldo Quevedo Cuenca María Laura Matos Mónica Hebe Vazquez-Levin |
| author_sort | Marina Rosso |
| collection | DOAJ |
| description | Abstract Background 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 done to characterize biological processes and canonical pathways affected by E-cadherin variant expression. 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. |
| first_indexed | 2024-12-10T05:22:32Z |
| format | Article |
| id | doaj.art-6fea8179c1ae4f2ebdd22a4d8f1d0f40 |
| institution | Directory Open Access Journal |
| issn | 2049-3002 |
| language | English |
| last_indexed | 2024-12-10T05:22:32Z |
| publishDate | 2019-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer & Metabolism |
| spelling | doaj.art-6fea8179c1ae4f2ebdd22a4d8f1d0f402022-12-22T02:00:46ZengBMCCancer & Metabolism2049-30022019-05-017112310.1186/s40170-019-0196-9Characterization 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 BMarina Rosso0Lara Lapyckyj1María José Besso2Marta Monge3Jaume Reventós4Francesc Canals5Jorge Oswaldo Quevedo Cuenca6María Laura Matos7Mónica Hebe Vazquez-Levin8Laboratorio de Estudios de Interacción Celular en Reproducción y Cáncer, Instituto de Biología y Medicina Experimental (IBYME; CONICET-FIBYME)Laboratorio de Estudios de Interacción Celular en Reproducción y Cáncer, Instituto de Biología y Medicina Experimental (IBYME; CONICET-FIBYME)Laboratorio de Estudios de Interacción Celular en Reproducción y Cáncer, Instituto de Biología y Medicina Experimental (IBYME; CONICET-FIBYME)Vall d’Hebron Institute of Oncology (VHIO)Departament de Ciències Bàsiques, Universitat Internacional de CatalunyaVall d’Hebron Institute of Oncology (VHIO)Laboratorio de Estudios de Interacción Celular en Reproducción y Cáncer, Instituto de Biología y Medicina Experimental (IBYME; CONICET-FIBYME)Laboratorio de Estudios de Interacción Celular en Reproducción y Cáncer, Instituto de Biología y Medicina Experimental (IBYME; CONICET-FIBYME)Laboratorio de Estudios de Interacción Celular en Reproducción y Cáncer, Instituto de Biología y Medicina Experimental (IBYME; CONICET-FIBYME)Abstract Background 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 done to characterize biological processes and canonical pathways affected by E-cadherin variant expression. 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.http://link.springer.com/article/10.1186/s40170-019-0196-9Breast cancerEpithelial cadherinAlternative splicingEpithelial to mesenchymal transitionProteomic analysis2D-DIGE |
| spellingShingle | Marina Rosso Lara Lapyckyj María José Besso Marta Monge Jaume Reventós Francesc Canals Jorge Oswaldo Quevedo Cuenca María Laura Matos Mónica Hebe Vazquez-Levin 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 Cancer & Metabolism Breast cancer Epithelial cadherin Alternative splicing Epithelial to mesenchymal transition Proteomic analysis 2D-DIGE |
| 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 |
| 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_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_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 |
| topic | Breast cancer Epithelial cadherin Alternative splicing Epithelial to mesenchymal transition Proteomic analysis 2D-DIGE |
| url | http://link.springer.com/article/10.1186/s40170-019-0196-9 |
| work_keys_str_mv | AT marinarosso characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT laralapyckyj characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT mariajosebesso characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT martamonge characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT jaumereventos characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT francesccanals characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT jorgeoswaldoquevedocuenca characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT marialauramatos characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen AT monicahebevazquezlevin characterizationofthemolecularchangesassociatedwiththeoverexpressionofanovelepithelialcadherinsplicevariantmrnainabreastcancermodelusingproteomicsandbioinformaticsapproachesidentificationofchangesincellmetabolismandanincreasedexpressionoflactatedehydrogen |