title: Proteomic Analysis of Exosomes and Exosome-Free Conditioned Media From Human Osteosarcoma Cell Lines Reveals Secretion of Proteins Related to Tumor Progression.
abstract: Osteosarcomas are the most prevalent bone tumors in pediatric patients, but can also occur later in life. Bone tumors have the potential to metastasize to lung and occasionally other vital organs. To understand how osteosarcoma cells interact with their micro-environment to support bone tumor progression and metastasis, we analyzed secreted proteins and exosomes from three human osteosarcoma cell lines. Exosome isolation was validated by transmission electron microscopy (TEM) and immuno-blotting for characteristic biomarkers (CD63, CD9, and CD81). Exosomal and soluble proteins (less than 100 kDa) were identified by mass spectrometry analysis using nanoLC-MS/MS and classified by functional gene ontology clustering. We identified a secretome set of >3,000 proteins for both fractions, and detected proteins that are either common or unique among the three osteosarcoma cell lines. Protein ontology comparison of proteomes from exosomes and exosome-free fractions revealed differences in the enrichment of functional categories associated with different biological processes, including those related to tumor progression (i.e., angiogenesis, cell adhesion, and cell migration). The secretome characteristics of osteosarcoma cells are consistent with the pathological properties of tumor cells with metastatic potential. J. Cell. Biochem. 118: 351-360, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Millennium Institute on Immunology and Immunotherapy, University of Chile, Santiago, Chile.|Program of Cellular and Molecular Biology, Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, Santiago, Chile.
authors.forename: Sofía
authors.initials: S
authors.lastname: Jerez
authors.affiliation: Millennium Institute on Immunology and Immunotherapy, University of Chile, Santiago, Chile.|Program of Cellular and Molecular Biology, Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, Santiago, Chile.
authors.forename: Héctor
authors.initials: H
authors.lastname: Araya
authors.affiliation: Departments of Orthopedic Surgery & Biochemistry and Molecular Biology, Mayo Clinic, 200 First Street S.W., MSB 3-69, Rochester, Minnesota 55905.
authors.forename: Roman
authors.initials: R
authors.lastname: Thaler
authors.affiliation: Mayo Proteomics Research Center, Mayo Clinic, Rochester, Minnesota 55905.
authors.forename: M Cristine
authors.initials: MC
authors.lastname: Charlesworth
authors.affiliation: Program of Cellular and Molecular Biology, Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, Santiago, Chile.
authors.forename: Remigio
authors.initials: R
authors.lastname: López-Solís
authors.affiliation: Millennium Institute on Immunology and Immunotherapy, Departamento de Genética Molecular y Microbiología Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.|Departamento de Endocrinología, Facultad de Medicina, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Alexis M
authors.initials: AM
authors.lastname: Kalergis
authors.affiliation: Millennium Institute on Immunology and Immunotherapy, Departamento de Genética Molecular y Microbiología Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Pablo F
authors.initials: PF
authors.lastname: Céspedes
authors.affiliation: Departments of Orthopedic Surgery & Biochemistry and Molecular Biology, Mayo Clinic, 200 First Street S.W., MSB 3-69, Rochester, Minnesota 55905.
authors.forename: Amel
authors.initials: A
authors.lastname: Dudakovic
authors.affiliation: Department of Biochemistry, University of Vermont College of Medicine, 89 Beaumont Avenue, Burlington, Vermont 05405.
authors.forename: Gary S
authors.initials: GS
authors.lastname: Stein
authors.affiliation: Departments of Orthopedic Surgery & Biochemistry and Molecular Biology, Mayo Clinic, 200 First Street S.W., MSB 3-69, Rochester, Minnesota 55905.
authors.forename: Andre J
authors.initials: AJ
authors.lastname: van Wijnen
authors.affiliation: Millennium Institute on Immunology and Immunotherapy, University of Chile, Santiago, Chile.|Program of Cellular and Molecular Biology, Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, Santiago, Chile.
authors.forename: Mario
authors.initials: M
authors.lastname: Galindo
mesh_terms: D015703:Antigens, CD / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001859:Bone Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D055354:Exosomes / Q000378:metabolism*; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*; D012516:Osteosarcoma / Q000378:metabolism*
chemical_list: D015703:Antigens, CD; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins
keywords: CANCER; EXOSOME; OSTEOSARCOMA; SECRETOME; TUMOR PROGRESSION