Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines

Rationale: Gastric cancer (GC) is a solid tumor that contains subpopulations of cancer stem cells (CSCs), which are considered drivers of tumor initiation and metastasis; responsible for therapeutic resistance; and promoters of tumor relapse. The balance between symmetric and asymmetric division is...

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Authors: Pajuelo-Lozano, Natalia, Alcalá, Sonia, Sainz, Bruno, Perona, Rosario, Sánchez Pérez, María Isabel
Format: article
Publication Date:2020
Country:España
Institution:Universidad Autónoma de Madrid
Repository:Biblos-e Archivo. Repositorio Institucional de la UAM
Language:English
OAI Identifier:oai:repositorio.uam.es:10486/695186
Online Access:http://hdl.handle.net/10486/695186
https://dx.doi.org/10.7150/thno.49270
Access Level:Open access
Keyword:EMT
Gastric cancer stem-like cells
MAD2
MMPs
Tumorigenesis
Medicina
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spelling Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell linesPajuelo-Lozano, NataliaAlcalá, SoniaSainz, BrunoPerona, RosarioSánchez Pérez, María IsabelEMTGastric cancer stem-like cellsMAD2MMPsTumorigenesisMedicinaRationale: Gastric cancer (GC) is a solid tumor that contains subpopulations of cancer stem cells (CSCs), which are considered drivers of tumor initiation and metastasis; responsible for therapeutic resistance; and promoters of tumor relapse. The balance between symmetric and asymmetric division is crucial for stem cell maintenance. The objective of this study is to evaluate the role of MAD2, a key protein for proper mitotic checkpoint activity, in the tumorigenesis of GC. Methods: Gastric cancer stem cells (GCSCs) were obtained from MKN45, SNU638 and ST2957 cell lines. Pluripotency and stemness markers were evaluated by RT-qPCR and autofluorescence and membrane markers by flow cytometry. Relevant signal transduction pathways were studied by WB. We analysed cell cycle progression, migration and invasion after modulation of MAD2 activity or protein expression levels in these in vitro models. In vivo assays were performed in a nude mouse subcutaneous xenograft model. Results: We found that NANOG, CXCR4 and autofluorescence are common and consistent markers for the GCSCs analysed, with other markers showing more variability. The three main signalling pathways (Wnt/β-catenin; Hedgehog and Notch) were activated in GCSCs. Downregulation of MAD2 in MKN45CSCs decreased the expression of markers CXCR4, CD133, CD90, LGR5 and VIM, without affecting cell cycle profile or therapy resistance. Moreover, migration, invasion and tumor growth were clearly reduced, and accordingly, we found that metalloprotease expression decreased. These results were accompanied by a reduction in the levels of transcription factors related with epithelial-to-mesenchymal transition. Conclusions: We can conclude that MAD2 is important for GCSCs stemness and its downregulation in MKN45CSCs plays a central role in GC tumorigenesis, likely through CXCR4-SNAI2-MMP1. Thus, its potential use in the clinical setting should be studied as its functions appear to extend beyond mitosis.This work was supported by Fondo de Investigaciones Sanitarias (FIS), Instituto de Salud Carlos III, Spain, grants PI17-01401 (PR) and PI18/00757 (BS,Jr), (all co-financed through Fondo Europeo de Desarrollo Regional (FEDER) “Una manera de hacer Europa”). NPL was supported by a grant FPU15/04669, funded by Ministerio de Educación, Cultura y Deporte, Spain. NPL is a fellow of the Doctoral Program in Molecular Biosciences, UAM, Madrid, Spain. BS,Jr was supported by a Ramón y Cajal Merit Award (RYC-2012-12104) from the Ministerio de Economía y Competitividad, SpainIvyspring International PublisherDepartamento de BioquímicaFacultad de MedicinaInstituto de Investigaciones Biomédicas "Alberto Sols" (IIBM)Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS)20202020-06-25research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10486/695186https://dx.doi.org/10.7150/thno.49270reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/6951862026-06-23T12:46:27Z
dc.title.none.fl_str_mv Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
title Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
spellingShingle Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
Pajuelo-Lozano, Natalia
EMT
Gastric cancer stem-like cells
MAD2
MMPs
Tumorigenesis
Medicina
title_short Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
title_full Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
title_fullStr Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
title_full_unstemmed Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
title_sort Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
dc.creator.none.fl_str_mv Pajuelo-Lozano, Natalia
Alcalá, Sonia
Sainz, Bruno
Perona, Rosario
Sánchez Pérez, María Isabel
author Pajuelo-Lozano, Natalia
author_facet Pajuelo-Lozano, Natalia
Alcalá, Sonia
Sainz, Bruno
Perona, Rosario
Sánchez Pérez, María Isabel
author_role author
author2 Alcalá, Sonia
Sainz, Bruno
Perona, Rosario
Sánchez Pérez, María Isabel
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Departamento de Bioquímica
Facultad de Medicina
Instituto de Investigaciones Biomédicas "Alberto Sols" (IIBM)
Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS)
dc.subject.none.fl_str_mv EMT
Gastric cancer stem-like cells
MAD2
MMPs
Tumorigenesis
Medicina
topic EMT
Gastric cancer stem-like cells
MAD2
MMPs
Tumorigenesis
Medicina
description Rationale: Gastric cancer (GC) is a solid tumor that contains subpopulations of cancer stem cells (CSCs), which are considered drivers of tumor initiation and metastasis; responsible for therapeutic resistance; and promoters of tumor relapse. The balance between symmetric and asymmetric division is crucial for stem cell maintenance. The objective of this study is to evaluate the role of MAD2, a key protein for proper mitotic checkpoint activity, in the tumorigenesis of GC. Methods: Gastric cancer stem cells (GCSCs) were obtained from MKN45, SNU638 and ST2957 cell lines. Pluripotency and stemness markers were evaluated by RT-qPCR and autofluorescence and membrane markers by flow cytometry. Relevant signal transduction pathways were studied by WB. We analysed cell cycle progression, migration and invasion after modulation of MAD2 activity or protein expression levels in these in vitro models. In vivo assays were performed in a nude mouse subcutaneous xenograft model. Results: We found that NANOG, CXCR4 and autofluorescence are common and consistent markers for the GCSCs analysed, with other markers showing more variability. The three main signalling pathways (Wnt/β-catenin; Hedgehog and Notch) were activated in GCSCs. Downregulation of MAD2 in MKN45CSCs decreased the expression of markers CXCR4, CD133, CD90, LGR5 and VIM, without affecting cell cycle profile or therapy resistance. Moreover, migration, invasion and tumor growth were clearly reduced, and accordingly, we found that metalloprotease expression decreased. These results were accompanied by a reduction in the levels of transcription factors related with epithelial-to-mesenchymal transition. Conclusions: We can conclude that MAD2 is important for GCSCs stemness and its downregulation in MKN45CSCs plays a central role in GC tumorigenesis, likely through CXCR4-SNAI2-MMP1. Thus, its potential use in the clinical setting should be studied as its functions appear to extend beyond mitosis.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-06-25
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10486/695186
https://dx.doi.org/10.7150/thno.49270
url http://hdl.handle.net/10486/695186
https://dx.doi.org/10.7150/thno.49270
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Ivyspring International Publisher
publisher.none.fl_str_mv Ivyspring International Publisher
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
repository.name.fl_str_mv
repository.mail.fl_str_mv
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