Molecular and Cellular Biochemistry

, Volume 275, Issue 1–2, pp 25–55 | Cite as

Proteome profiling of human epithelial ovarian cancer cell line TOV-112D

  • Jean-Philippe Gagné
  • Pierre Gagné
  • Joanna M. Hunter
  • Marie-Ève Bonicalzi
  • Jean-François Lemay
  • Isabelle Kelly
  • Cécile Le Page
  • Diane Provencher
  • Anne-Marie Mes-Masson
  • Arnaud Droit
  • David Bourgais
  • Guy G. Poirier


A proteome profiling of the epithelial ovarian cancer cell line TOV-112D was initiated as a protein expression reference in the study of ovarian cancer. Two complementary proteomic approaches were used in order to maximise protein identification: two-dimensional gel electrophoresis (2DE) protein separation coupled to matrix assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and one-dimensional gel electrophoresis (1DE) coupled to liquid-chromatography tandem mass spectrometry (LC MS/MS). One hundred and seventy-two proteins have been identified among 288 spots selected on two-dimensional gels and a total of 579 proteins were identified with the 1DE LC MS/MS approach. This proteome profiling covers a wide range of protein expression and identifies several proteins known for their oncogenic properties. Bioinformatics tools were used to mine databases in order to determine whether the identified proteins have previously been implicated in pathways associated with carcinogenesis or cell proliferation. Indeed, several of the proteins have been reported to be specific ovarian cancer markers while others are common to many tumorigenic tissues or proliferating cells. The diversity of proteins found and their association with known oncogenic pathways validate this proteomic approach. The proteome 2D map of the TOV-112D cell line will provide a valuable resource in studies on differential protein expression of human ovarian carcinomas while the 1DE LC MS/MS approach gives a picture of the actual protein profile of the TOV-112D cell line. This work represents one of the most complete ovarian protein expression analysis reports to date and the first comparative study of gene expression profiling and proteomic patterns in ovarian cancer.


2D-gel mass spectrometry protein expression transcriptome 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Whitman G, Nolan T, Gallup D: Normal ovarian development and function. In: M. Markman, W.J. Hoskins (eds). Cancer of the Ovary. Raven Press, New York, 1993Google Scholar
  2. 2.
    Chang J, Fryatt I, Ponder B, Fisher C, Gore ME: A matched control study of familial epithelial ovarian cancer: Patient characteristics, response to chemotherapy and outcome. Ann Oncol 6: 80–82, 1995Google Scholar
  3. 3.
    Kaku T, Ogawa S, Kawano Y, Ohishi Y, Kobayashi H, Hirakawa T, Nakano H: Histological classification of ovarian cancer. Med Electron Microsc 36: 9–17, 2003Google Scholar
  4. 4.
    Provencher DM, Lounis H, Champoux L, Tetrault M, Manderson EN, Wang JC, Eydoux P, Savoie R, Tonin PN, Mes-Masson AM: Characterization of four novel epithelial ovarian cancer cell lines. In Vitro Cell Dev Biol Anim 36: 357–361, 2000Google Scholar
  5. 5.
    Keller A, Nesvizhskii AI, Kolker E, Aebersold R: Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem 74: 5383–5392, 2002Google Scholar
  6. 6.
    Von Haller PD, Yi E, Donohoe S, Vaughn K, Keller A, Nesvizhskii AI, Eng J, Li XJ, Goodlett DR, Aebersold R, Watts JD: The application of new software tools to quantitative protein profiling via Isotope-coded Affinity Tag (ICAT) and Tandem Mass Spectrometry: II. Evaluation of Tandem Mass Spectrometry Methodologies for Large-Scale Protein Analysis, and the Application of Statistical Tools for Data Analysis and Interpretation. Mol Cell Proteomics 2: 428–442, 2003Google Scholar
  7. 7.
    Nesvizhskii AI, Keller A, Kolker E, Aebersold R: A statistical model for identifying proteins by tandem mass spectrometry. Anal Chem 75: 4646–4658, 2003Google Scholar
  8. 8.
    Conroy SE, Latchman DS: Do heat shock proteins have a role in breast cancer? Br J Cancer 74: 717–721, 1996Google Scholar
  9. 9.
    Le Page C, Provencher D, Maugard CM, Ouellet V, Mes-Masson AM: Signature of a silent killer: Expression profiling in epithelial ovarian cancer. Expert Rev Mol Diagn 4: 157–167, 2004Google Scholar
  10. 10.
    Tonin PN, Hudson TJ, Rodier F, Bossolasco M, Lee PD, Novak J, Manderson EN, Provencher D, Mes-Masson AM: Microarray analysis of gene expression mirrors the biology of an ovarian cancer model. Oncogene 20: 6617–6626, 2001Google Scholar
  11. 11.
    Sladek NE, Kollander R, Sreerama L, Kiang DT: Cellular levels of aldehyde dehydrogenases (ALDH1A1 and ALDH3A1) as predictors of therapeutic responses to cyclophosphamide-based chemotherapy of breast cancer: A retrospective study. Rational individualization of oxazaphosphorine-based cancer chemotherapeutic regimens. Cancer Chemother Pharmacol 49: 309–321, 2002Google Scholar
  12. 12.
    Bergman AC, Benjamin T, Alaiya A, Waltham M, Sakaguchi K, Franzen B, Linder S, Bergman T, Auer G, Appella E, Wirth PJ, Jornvall H: Identification of gel-separated tumor marker proteins by mass spectrometry. Electrophoresis 21: 679–686, 2000Google Scholar
  13. 13.
    Altenberg B, Greulich KO: Genes of glycolysis are ubiquitously overexpressed in 24 cancer classes. Genomics 84: 1014–1020, 2004Google Scholar
  14. 14.
    Jones MB, Krutzsch H, Shu H, Zhao Y, Liotta LA, Kohn EC, Petricoin EF, 3rd: Proteomic analysis and identification of new biomarkers and therapeutic targets for invasive ovarian cancer. Proteomics 2: 76–84, 2002Google Scholar
  15. 15.
    Alaiya AA, Franzen B, Fujioka K, Moberger B, Schedvins K, Silfversvard C, Linder S, Auer G: Phenotypic analysis of ovarian carcinoma: Polypeptide expression in benign, borderline and malignant tumors. Int J Cancer 73: 678–683, 1997Google Scholar
  16. 16.
    Tomic S, Ilic Forko J, Babic D, Sundov D, Kuret S, Andelinovic S: c-erbB-2, p53, and nm23 proteins as prognostic factors in patients with epithelial ovarian carcinoma. Croat Med J 44: 429–434, 2003Google Scholar
  17. 17.
    Simone G, Falco G, Caponio MA, Campobasso C, De Frenza M, Petroni S, Wiesel S, Leone A: nm23 expression in malignant ascitic effusions of serous ovarian adenocarcinoma. Int J Oncol 19: 885–890, 2001Google Scholar
  18. 18.
    Schneider J, Pollan M, Jimenez E, Marenbach K, Martinez N, Volm M, Marx D, Meden H: nm23-H1 expression defines a high-risk subpopulation of patients with early-stage epithelial ovarian carcinoma. Br J Cancer 82: 1662–1670, 2000Google Scholar
  19. 19.
    Qian M, Feng Y, Xu L, Zheng S, Zhou X: Expression of antimetastatic gene nm23-H1 in epithelial ovarian cancer. Chin Med J (Engl) 110: 142–144, 1997Google Scholar
  20. 20.
    Stierum R, Gaspari M, Dommels Y, Ouatas T, Pluk H, Jespersen S, Vogels J, Verhoeckx K, Groten J, van Ommen B: Proteome analysis reveals novel proteins associated with proliferation and differentiation of the colorectal cancer cell line Caco-2. Biochim Biophys Acta 1650: 73–91, 2003Google Scholar
  21. 21.
    Yow HK, Wong JM, Chen HS, Lee CG, Davis S, Steele GD, Jr., Chen LB: Increased mRNA expression of a laminin-binding protein in human colon carcinoma: Complete sequence of a full-length cDNA encoding the protein. Proc Natl Acad Sci USA 85: 6394–6398, 1988Google Scholar
  22. 22.
    Mathur S, Cleary KR, Inamdar N, Kim YH, Steck P, Frazier ML: Overexpression of elongation factor-1gamma protein in colorectal carcinoma. Cancer 82: 816–821, 1998Google Scholar
  23. 23.
    Zhang Y, Woodford N, Xia X, Hamburger AW: Repression of E2F1-mediated transcription by the ErbB3 binding protein Ebp1 involves histone deacetylases. Nucleic Acids Res 31: 2168–2177, 2003Google Scholar
  24. 24.
    Hirst M, Haliday E, Nakamura J, Lou L: Human GMP synthetase. Protein purification, cloning, and functional expression of cDNA. J Biol Chem 269: 23830–23837, 1994Google Scholar
  25. 25.
    Wells J, Henkler F, Leversha M, Koshy R: A mitochondrial elongation factor-like protein is over-expressed in tumours and differentially expressed in normal tissues. FEBS Lett 358: 119–125, 1995Google Scholar
  26. 26.
    Fukuda S, Wu DW, Stark K, Pelus LM: Cloning and characterization of a proliferation-associated cytokine-inducible protein, CIP29. Biochem Biophys Res Commun 292: 593–600, 2002Google Scholar
  27. 27.
    Fujii J, Ikeda Y: Advances in our understanding of peroxiredoxin, a multifunctional, mammalian redox protein. Redox Rep 7: 123–130, 2002Google Scholar
  28. 28.
    Yanai Y, Micallef MJ, Yamamoto S, Yamamoto K, Yamauchi H, Ikegami H, Kurimoto M: Expression profiling of tumor necrosis factor alpha-induced apoptosis-associated genes in human solid tumor cell lines. Anticancer Res 23: 2339–2348, 2003Google Scholar
  29. 29.
    Rosenquist M: 14-3-3 proteins in apoptosis. Braz J Med Biol Res 36: 403–408, 2003Google Scholar
  30. 30.
    Weis K: Regulating access to the genome: Nucleocytoplasmic transport throughout the cell cycle. Cell 112: 441–451, 2003Google Scholar
  31. 31.
    Rosenfeld JL, Knoll BJ, Moore RH: Regulation of G-protein-coupled receptor activity by rab GTPases. Receptors Channels 8: 87–97, 2002Google Scholar
  32. 32.
    McClung JK, King RL, Walker LS, Danner DB, Nuell MJ, Stewart CA, Dell’Orco RT: Expression of prohibitin, an antiproliferative protein. Exp Gerontol 27: 413–417, 1992Google Scholar
  33. 33.
    Wang S, Nath N, Adlam M, Chellappan S: Prohibitin, a potential tumor suppressor, interacts with RB and regulates E2F function. Oncogene 18: 3501–3510, 1999Google Scholar
  34. 34.
    Jupe ER, Liu XT, Kiehlbauch JL, McClung JK, Dell’Orco RT: Prohibitin in breast cancer cell lines: Loss of antiproliferative activity is linked to 3′ untranslated region mutations. Cell Growth Differ 7: 871–878, 1996Google Scholar
  35. 35.
    Wu Y, Pan S, Che S, He G, Nelman-Gonzalez M, Weil MM, Kuang J: Overexpression of Hp95 induces G1 phase arrest in confluent HeLa cells. Differentiation 67: 139–153, 2001Google Scholar
  36. 36.
    Leonard GD, Low JA, Berman AW, Swain SM: CA 125 elevation in breast cancer: A case report and review of the literature. Breast J 10: 146–149, 2004Google Scholar
  37. 37.
    Sun HT, Cohen S, Kaufmann WE: Annexin-1 is abnormally expressed in fragile X syndrome: Two-dimensional electrophoresis study in lymphocytes. Am J Med Genet 103: 81–90, 2001Google Scholar
  38. 38.
    Oh P, Li Y, Yu J, Durr E, Krasinska KM, Carver LA, Testa JE, Schnitzer JE: Subtractive proteomic mapping of the endothelial surface in lung and solid tumours for tissue-specific therapy. Nature 429: 629–635, 2004Google Scholar
  39. 39.
    Eng J, McCormack A, Yates JR, III: An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database. J Am Soc Mass Spectrom 5: 976–989, 1994Google Scholar
  40. 40.
    Tamayo P, Slonim D, Mesirov J, Zhu Q, Kitareewan S, Dmitrovsky E, Lander ES, Golub TR: Interpreting patterns of gene expression with self-organizing maps: Methods and application to hematopoietic differentiation. Proc Natl Acad Sci USA 96: 2907–2912, 1999Google Scholar
  41. 41.
    Novak P, Sladek R, Hudson TJ: Characterization of variability in large-scale gene expression data: Implications for study design. Genomics 79: 104–113, 2002Google Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Jean-Philippe Gagné
    • 1
  • Pierre Gagné
    • 1
  • Joanna M. Hunter
    • 1
  • Marie-Ève Bonicalzi
    • 1
  • Jean-François Lemay
    • 2
  • Isabelle Kelly
    • 2
  • Cécile Le Page
    • 3
  • Diane Provencher
    • 3
  • Anne-Marie Mes-Masson
    • 3
  • Arnaud Droit
    • 2
  • David Bourgais
    • 2
  • Guy G. Poirier
    • 1
    • 2
    • 4
  1. 1.Health and Environment UnitLaval University Medical Research Center, CHUQ, Faculty of Medicine, Laval UniversityCanada
  2. 2.Eastern Québec Proteomic Center, CHUQCanada
  3. 3.Centre de recherche du CentreHospitalier de l’Université de Montréal (CHUM)-Hôpital Notre–Dame and Institut du cancer de MontréalMontréalCanada
  4. 4.Health and Environment Unit/Eastern Québec Proteomic CenterLaval University Medical Research Center, CHUQ, Faculty of Medicine, Laval UniversityCanada

Personalised recommendations