Targeting MAD2 modulates stemness and tumorigenesis in human gastric cancer cell lines
EntityUAM. Departamento de Bioquímica; Instituto de Investigaciones Biomédicas "Alberto Sols" (IIBM); Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS)
PublisherIvyspring International Publisher
10.7150/thno.49270Theranostics 10.21 (2020): 9601-9618
Funded byThis 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, Spain
ProjectGobierno de España. PI17-01401; Gobierno de España. PI18/00757; Gobierno de España. RYC-2012-12104
SubjectsEMT; Gastric cancer stem-like cells; MAD2; MMPs; Tumorigenesis; Medicina
Rights© 2020 The Authors
Esta obra está bajo una Licencia Creative Commons Atribución 4.0 Internacional.
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.
Google Scholar:Pajuelo-Lozano, Natalia - Alcalá, Sonia - Sainz, Bruno - Perona, Rosario - Sanchez-Perez, Isabel
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