title: DNA methylation profiling identifies PTRF/Cavin-1 as a novel tumor suppressor in Ewing sarcoma when co-expressed with caveolin-1.
abstract: Epigenetic modifications have been shown to be important in developmental tumors as Ewing sarcoma. We profiled the DNA methylation status of 15 primary tumors, 7 cell lines, 10 healthy tissues and 4 human mesenchymal stem cells lines samples using the Infinium Human Methylation 450K. Differential methylation analysis between Ewing sarcoma and reference samples revealed 1166 hypermethylated and 864 hypomethylated CpG sites (Bonferroni p < 0.05, δ-β-value with absolute difference of >0.20) corresponding to 392 and 470 genes respectively. Gene Ontology analysis of genes differentially methylated in Ewing sarcoma samples showed a significant enrichment of developmental genes. Membrane and cell signal genes were also enriched, among those, 11 were related to caveola formation. We identified differential hypermethylation of CpGs located in the body and S-Shore of the PTRF gene in Ewing sarcoma that correlated with its repressed transcriptional state. Reintroduction of PTRF/Cavin-1 in Ewing sarcoma cells revealed a role of this protein as a tumor suppressor. Restoration of caveolae in the membrane of Ewing sarcoma cells, by exogenously reintroducing PTRF, disrupts the MDM2/p53 complex, which consequently results in the activation of p53 and the induction of apoptosis.
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Huertas-Martínez
authors.affiliation: Cancer Epigenetics and Biology Programme (PEBC), Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Franck
authors.initials: F
authors.lastname: Court
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Rello-Varona
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Herrero-Martín
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Olga
authors.initials: O
authors.lastname: Almacellas-Rabaiget
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Sáinz-Jaspeado
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Silvia
authors.initials: S
authors.lastname: Garcia-Monclús
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Lagares-Tena
authors.affiliation: Institut Germans Trias i Pujol (IGTP), Program in Predictive and Personalized Medicine of Cancer (IMPPC), Badalona, Barcelona, 08916, Spain.
authors.forename: Raquel
authors.initials: R
authors.lastname: Buj
authors.affiliation: Laboratory of Molecular Pathology, Instituto de Biomedicina de Sevilla (IBiS), Sevilla, 41013, Spain.
authors.forename: Lourdes
authors.initials: L
authors.lastname: Hontecillas-Prieto
authors.affiliation: Unidad de Hematología y Oncología Pediátrica, Hospital Universitario La Paz, Madrid, 28046, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Sastre
authors.affiliation: Servicio de Anatomía Patológica, Hospital Infantil Universitario Niño Jesús, Madrid, 28009, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Azorin
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain; Department of Pathology, Hospital Universitari de Bellvitge (HUB), L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Sanjuan
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Roser
authors.initials: R
authors.lastname: López-Alemany
authors.affiliation: Cancer Epigenetics and Biology Programme (PEBC), Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Moran
authors.affiliation: Laboratory of Translational Research in Pediatric Cancer, Vall d'Hebron Research Institute, Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, Barcelona, 08035, Spain.
authors.forename: Josep
authors.initials: J
authors.lastname: Roma
authors.affiliation: Laboratory of Translational Research in Pediatric Cancer, Vall d'Hebron Research Institute, Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, Barcelona, 08035, Spain.
authors.forename: Soledad
authors.initials: S
authors.lastname: Gallego
authors.affiliation: Developmental Tumor Biology Laboratory, Hospital Sant Joan de Deu, Esplugues de Llobregat, Barcelona, 08950, Spain.
authors.forename: Jaume
authors.initials: J
authors.lastname: Mora
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain; Sarcoma Multidisciplinary Unit, Institut Català d'Oncologia-ICO, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: García Del Muro
authors.affiliation: Department of Internal Medicine, University of Iowa, Iowa City, IA, 52242, USA.
authors.forename: Paloma H
authors.initials: PH
authors.lastname: Giangrande
authors.affiliation: Institut Germans Trias i Pujol (IGTP), Program in Predictive and Personalized Medicine of Cancer (IMPPC), Badalona, Barcelona, 08916, Spain.
authors.forename: Miquel A
authors.initials: MA
authors.lastname: Peinado
authors.affiliation: Unidad de Tumores Sólidos Infantiles, Instituto de Investigación de Enfermedades Raras, ISCIII, Madrid, 28029, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Alonso
authors.affiliation: Laboratory of Molecular Pathology, Instituto de Biomedicina de Sevilla (IBiS), Sevilla, 41013, Spain; Hospital Universitario Virgen del Rocio/CSIC/Universidad de Sevilla, Sevilla, 41013, Spain.
authors.forename: Enrique
authors.initials: E
authors.lastname: de Alava
authors.affiliation: Cancer Epigenetics and Biology Programme (PEBC), Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain.
authors.forename: Dave
authors.initials: D
authors.lastname: Monk
authors.affiliation: Cancer Epigenetics and Biology Programme (PEBC), Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain; Department of Physiological Sciences II, School of Medicine, University of Barcelona, Barcelona, Catalonia, Spain; Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Catalonia, Spain.
authors.forename: Manel
authors.initials: M
authors.lastname: Esteller
authors.affiliation: Sarcoma Research Group, Institut d'Investigació Biomèdica de Bellvitge-IDIBELL, L'Hospitalet de Llobregat, Barcelona, 08908, Spain. Electronic address: omartinez@idibell.cat.
authors.forename: Oscar M
authors.initials: OM
authors.lastname: Tirado
mesh_terms: D000818:Animals; D017209:Apoptosis; D001859:Bone Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D051242:Caveolin 1 / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D016147:Genes, Tumor Suppressor; D006801:Humans; D053208:Kaplan-Meier Estimate; D008819:Mice, Nude; D010766:Phosphorylation; D051736:Proto-Oncogene Proteins c-mdm2 / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D012512:Sarcoma, Ewing / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015398:Signal Transduction; D013030:Spain; D014162:Transfection; D047368:Tumor Burden; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: C495540:CAV1 protein, human; C497835:CAVIN1 protein, human; D051242:Caveolin 1; D016601:RNA-Binding Proteins; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2
keywords: Apoptosis; Caveolae; Cavin-1; DNA methylation; Ewing sarcoma; PTRF