The proteome profile of the human osteosarcoma U2OS cell line.

The human osteosarcoma U2OS cell line is one of the first generated cell lines and is used in various areas of biomedical research. Knowledge of its protein expression is limited and no comprehensive study on the proteome of this cell line has been reported to date. Proteomics technology was used in order to analyse the proteins of the U2OS cell line. Total protein extracts were separated by two-dimensional gel electrophoresis (2-DE) and analysed by matrix-assisted laser desorption ionisation-mass spectrometry (MALDI-MS) and MALDI--MS-MS following in-gel digestion with trypsin and, finally, protein identification was carried out by peptide mass fingerprint (PMF) and post source decay (PSD), respectively. Approximately 3,000 spots were excised from two 2-DE gels and were analysed, resulting in the identification of 237 different gene products. The majority of the identified proteins were enzymes, regulatory proteins and RNA-associated proteins, while leukocyte markers and oncogenes were also present. Our findings include 11 protooncogenes (FKBP4, SRC8, PSD10, FUBP1, PARK7, NPM, PDIA1, OXRP, SET, TCTP and GRP75) related to the cancerous state of the U2OS cell line. The U2OS 2-DE database provides the basis for future protein studies.

[1]  Xiaoling Zhang,et al.  Hypoxia-induced Nucleophosmin Protects Cell Death through Inhibition of p53* , 2004, Journal of Biological Chemistry.

[2]  Harold L. Moses,et al.  STRAP and Smad7 Synergize in the Inhibition of Transforming Growth Factor β Signaling , 2000, Molecular and Cellular Biology.

[3]  B. Fox,et al.  The promising future of proteomics in cancer diagnosis and treatment , 2005, European journal of gastroenterology & hepatology.

[4]  Konstantinos Vougas,et al.  The Protein Profile of the Human Immature T-cell Line CCRF-CEM. , 2005, Cancer genomics & proteomics.

[5]  P. Cassoni,et al.  Constitutive and ligand-induced nuclear localization of oxytocin receptor , 2007, Journal of cellular and molecular medicine.

[6]  J. Hale,et al.  Application of proteomics for discovery of protein biomarkers. , 2003, Briefings in functional genomics & proteomics.

[7]  D. Levens,et al.  Loss of FBP function arrests cellular proliferation and extinguishes c‐myc expression , 2000, The EMBO journal.

[8]  Michelle D. Garrett,et al.  Cell cycle control and cancer. , 2001 .

[9]  Liang Zhu.  Tumour suppressor retinoblastoma protein Rb: a transcriptional regulator. , 2005, European journal of cancer.

[10]  J. Węsierska‐Gądek,et al.  The subcellular distribution of the p53 tumour suppressor, and organismal ageing. , 2005, Cellular & molecular biology letters.

[11]  A. Görg,et al.  Current two‐dimensional electrophoresis technology for proteomics , 2004, Proteomics.

[12]  D. Zimonjic,et al.  Amplification and overexpression of the EMS 1 oncogene, a possible prognostic marker, in human hepatocellular carcinoma. , 2003, The Journal of molecular diagnostics : JMD.

[13]  Jun Yokota,et al.  Sp1-Mediated Transcription of the Werner Helicase Gene Is Modulated by Rb and p53 , 1998, Molecular and Cellular Biology.

[14]  K. Cashman,et al.  The effect of conjugated linoleic acid on the viability and metabolism of human osteoblast-like cells. , 2005, Prostaglandins, leukotrienes, and essential fatty acids.

[15]  A. Nakagawara,et al.  Stabilization of p73 by nuclear IkappaB kinase-alpha mediates cisplatin-induced apoptosis. , 2007, Journal of Biological Chemistry.

[16]  A. Sandberg,et al.  Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors: osteosarcoma and related tumors. , 2003, Cancer genetics and cytogenetics.

[17]  Dimitris Kletsas,et al.  Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints , 2006, Nature.

[18]  Dimitris Kletsas,et al.  Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions , 2005, Nature.

[19]  Mario Plebani,et al.  Proteomics: the next revolution in laboratory medicine? , 2005, Clinica chimica acta; international journal of clinical chemistry.

[20]  M. Berger,et al.  Identification of potent anticancer activity in Ximenia americana aqueous extracts used by African traditional medicine. , 2006, Toxicology and applied pharmacology.

[21]  J. Healey,et al.  Cell Proliferation of Cultured Human Cancer Cells are Affected by the Elevated Tumor Pressures that Exist In Vivo , 2005, Annals of Biomedical Engineering.

[22]  B. Klein,et al.  Cell death in cultured human Saos2 osteoblasts exposed to low‐density lipoprotein , 2003, Journal of cellular biochemistry.

[23]  L. Hartwell,et al.  Cell cycle control and cancer. , 1994, Science.

[24]  Xiaoling Zhang,et al.  Negative regulation of p53 by nucleophosmin antagonizes stress-induced apoptosis in human normal and malignant hematopoietic cells. , 2005, Leukemia research.

[25]  P. Supaphol,et al.  Preparation and characterization of novel bone scaffolds based on electrospun polycaprolactone fibers filled with nanoparticles. , 2006, Macromolecular bioscience.

[26]  Xin Zhao,et al.  Interaction of DJ-1 with Daxx inhibits apoptosis signal-regulating kinase 1 activity and cell death. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Robert L. Sutherland,et al.  Cyclin D1, EMS1 and 11q13 Amplification in Breast Cancer , 2003, Breast Cancer Research and Treatment.

[28]  Hitoshi Suzuki,et al.  Thr199 phosphorylation targets nucleophosmin to nuclear speckles and represses pre‐mRNA processing , 2006, FEBS letters.

[29]  J. Squire,et al.  Chromosomal instability in osteosarcoma and its association with centrosome abnormalities. , 2003, Cancer genetics and cytogenetics.

[30]  P. Gregory,et al.  Isogenic Human Cell Lines for Drug Discovery: Regulation of Target Gene Expression by Engineered Zinc-Finger Protein Transcription Factors , 2005, Journal of biomolecular screening.

[31]  P. Sorensen,et al.  Spectral karyotyping identifies recurrent complex rearrangements of chromosomes 8, 17, and 20 in osteosarcomas , 2003, Genes, chromosomes & cancer.

[32]  R. Isfort,et al.  Analysis of oncogenes, tumor suppressor genes, autocrine growth‐factor production, and differentiation state of human osteosarcoma cell lines , 1995, Molecular carcinogenesis.

[33]  M. Paul-Clark,et al.  Calcitonin and prednisolone display antagonistic actions on bone and have synergistic effects in experimental arthritis. , 2007, The American journal of pathology.

[34]  G. Tsangaris,et al.  Protein profile of the HeLa cell line. , 2004, Journal of chromatography. A.

[35]  M. Buckle,et al.  Characterisation of the interface between nucleophosmin (NPM) and p53: Potential role in p53 stabilisation , 2006, FEBS letters.

[36]  P Berndt,et al.  Reliable automatic protein identification from matrix‐assisted laser desorption/ionization mass spectrometric peptide fingerprints , 1999, Electrophoresis.