Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study

In this work, the optimization of an effective protocol for cell metabolomics is described with special emphasis in the sample preparation and subsequent analysis of intracellular metabolites from adherent mammalian cells by capillary electrophoresis-mass spectrometry. As case study, colon cancer HT...

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Detalles Bibliográficos
Autores: Ibáñez, Clara, Simó, Carolina, Valdés, Alberto, Campone, Luca, Piccinelli, Anna Lisa, García-Cañas, Virginia, Cifuentes, Alejandro
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2015
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/113704
Acceso en línea:http://hdl.handle.net/10261/113704
Access Level:acceso abierto
Palabra clave:Human colon adenocarcinoma HT-29 cells
Metabolomics
Difluoromethylornithine
Polyamines
Capillary electrophoresis-mass spectrometry
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spelling Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case studyIbáñez, ClaraSimó, CarolinaSimó, CarolinaValdés, AlbertoCampone, LucaPiccinelli, Anna LisaGarcía-Cañas, VirginiaCifuentes, AlejandroHuman colon adenocarcinoma HT-29 cellsMetabolomicsDifluoromethylornithinePolyaminesCapillary electrophoresis-mass spectrometryIn this work, the optimization of an effective protocol for cell metabolomics is described with special emphasis in the sample preparation and subsequent analysis of intracellular metabolites from adherent mammalian cells by capillary electrophoresis-mass spectrometry. As case study, colon cancer HT-29 cells, a human cell model to investigate colon cancer, are employed. The feasibility of the whole method for cell metabolomics is demonstrated via a fast and sensitive profiling of the intracellular metabolites HT-29 cells by capillary electrophoresis-time-of-flight mass spectrometry (CE-TOF MS). The suitability of this methodology is further corroborated through the examination of the metabolic changes in the polyamines pathway produced in colon cancer HT-29 cells by difluoromethylornithine (DFMO), a known potent ornithine decarboxylase inhibitor. The selection of the optimum extraction conditions allowed a higher sample volume injection that led to an increase in CE-TOF MS sensitivity. Following a non-targeted metabolomics approach, 10 metabolites (namely, putrescine, ornithine, gamma-aminobutyric acid (GABA), oxidized and reduced glutathione, 5'-deoxy-5'-(methylthio)adenosine, N-acetylputrescine, cysteinyl-glycine, spermidine and an unknown compound) were found to be significantly altered by DFMO (p < 0.05) in HT-29 cells. In addition to the effect of DFMO on polyamine metabolism, minor modifications of other metabolic pathways (e.g., related to intracellular thiol redox state) were observed.This work was supported by an AGL2011-29857-C03-01 project (Ministerio de Educacion y Ciencia, Spain).Peer ReviewedElsevierMinisterio de Educación y Ciencia (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2015201520152015info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionhttp://hdl.handle.net/10261/113704reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1016/j.jpba.2015.03.001Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1137042026-05-22T06:33:51Z
dc.title.none.fl_str_mv Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
title Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
spellingShingle Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
Ibáñez, Clara
Human colon adenocarcinoma HT-29 cells
Metabolomics
Difluoromethylornithine
Polyamines
Capillary electrophoresis-mass spectrometry
title_short Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
title_full Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
title_fullStr Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
title_full_unstemmed Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
title_sort Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study
dc.creator.none.fl_str_mv Ibáñez, Clara
Simó, Carolina
Simó, Carolina
Valdés, Alberto
Campone, Luca
Piccinelli, Anna Lisa
García-Cañas, Virginia
Cifuentes, Alejandro
author Ibáñez, Clara
author_facet Ibáñez, Clara
Simó, Carolina
Valdés, Alberto
Campone, Luca
Piccinelli, Anna Lisa
García-Cañas, Virginia
Cifuentes, Alejandro
author_role author
author2 Simó, Carolina
Valdés, Alberto
Campone, Luca
Piccinelli, Anna Lisa
García-Cañas, Virginia
Cifuentes, Alejandro
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Educación y Ciencia (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Human colon adenocarcinoma HT-29 cells
Metabolomics
Difluoromethylornithine
Polyamines
Capillary electrophoresis-mass spectrometry
topic Human colon adenocarcinoma HT-29 cells
Metabolomics
Difluoromethylornithine
Polyamines
Capillary electrophoresis-mass spectrometry
description In this work, the optimization of an effective protocol for cell metabolomics is described with special emphasis in the sample preparation and subsequent analysis of intracellular metabolites from adherent mammalian cells by capillary electrophoresis-mass spectrometry. As case study, colon cancer HT-29 cells, a human cell model to investigate colon cancer, are employed. The feasibility of the whole method for cell metabolomics is demonstrated via a fast and sensitive profiling of the intracellular metabolites HT-29 cells by capillary electrophoresis-time-of-flight mass spectrometry (CE-TOF MS). The suitability of this methodology is further corroborated through the examination of the metabolic changes in the polyamines pathway produced in colon cancer HT-29 cells by difluoromethylornithine (DFMO), a known potent ornithine decarboxylase inhibitor. The selection of the optimum extraction conditions allowed a higher sample volume injection that led to an increase in CE-TOF MS sensitivity. Following a non-targeted metabolomics approach, 10 metabolites (namely, putrescine, ornithine, gamma-aminobutyric acid (GABA), oxidized and reduced glutathione, 5'-deoxy-5'-(methylthio)adenosine, N-acetylputrescine, cysteinyl-glycine, spermidine and an unknown compound) were found to be significantly altered by DFMO (p < 0.05) in HT-29 cells. In addition to the effect of DFMO on polyamine metabolism, minor modifications of other metabolic pathways (e.g., related to intracellular thiol redox state) were observed.
publishDate 2015
dc.date.none.fl_str_mv 2015
2015
2015
2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Postprint
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/113704
url http://hdl.handle.net/10261/113704
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1016/j.jpba.2015.03.001
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.811543

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