International Orthopaedics

, Volume 35, Issue 3, pp 401–411 | Cite as

Microarray analysis identifies distinct gene expression profiles associated with histological subtype in human osteosarcoma

  • Bernd Kubista
  • Florian Klinglmueller
  • Martin Bilban
  • Martin Pfeiffer
  • Richard Lass
  • Alexander Giurea
  • Phillipp T. Funovics
  • Cyril Toma
  • Martin Dominkus
  • Rainer Kotz
  • Theresia Thalhammer
  • Klemens Trieb
  • Teresa Zettl
  • Christian F. Singer
Original Paper

Abstract

Osteosarcoma is the most common primary malignant bone tumour. Currently osteosarcoma classification is based on histological appearance. It was the aim of this study to use a more systematic approach to osteosarcoma classification based on gene expression analysis and to identify subtype specific differentially expressed genes. We analysed the global gene expression profiles of ten osteosarcoma samples using Affymetrix U133A arrays (five osteoblastic and five non-osteoblastic osteosarcoma patients). Differential gene expression analysis yielded 75 genes up-regulated and 97 genes down-regulated in osteoblastic versus non-osteoblastic osteosarcoma samples, respectively. These included genes involved in cell growth, chemotherapy resistance, angiogenesis, steroid- and neuropeptide hormone receptor activity, acute-phase response and serotonin receptor activity and members of the Wnt/ß-catenin pathway and many others. Furthermore, we validated the highly differential expression of six genes including angiopoietin 1, IGFBP3, ferredoxin 1, BMP, decorin, and fibulin 1 in osteoblastic osteosarcoma relative to non-osteoblastic osteosarcoma. Our results show the utility of gene expression analysis to study osteosarcoma subtypes, and we identified several genes that may play a role as potential therapeutic targets in the future.

Notes

Conflicts of interest statement

None declared.

References

  1. 1.
    Nakano H, Tateishi A, Miki H et al (1999) Hyperthermic isolated regional perfusion for the treatment of osteosarcoma in the lower extremity. Am J Surg 178:27–32PubMedCrossRefGoogle Scholar
  2. 2.
    Davis AM, Bell RS, Goodwin PJ (1994) Prognostic factors in osteosarcoma: a critical review. J Clin Oncol 12:423–431PubMedGoogle Scholar
  3. 3.
    Ferrari S, Bertoni F, Mercuri M et al (2001) Predictive factors of disease-free survival for non-metastatic osteosarcoma of the extremity: an analysis of 300 patients treated at the Rizzoli Institute. Ann Oncol 12:1145–1150PubMedCrossRefGoogle Scholar
  4. 4.
    Hudson M, Jaffe MR, Jaffe N et al (1990) Pediatric osteosarcoma: therapeutic strategies, results, and prognostic factors derived from a 10-year experience. J Clin Oncol 8:1988–1997PubMedGoogle Scholar
  5. 5.
    Petrilli AS, Gentil FC, Epelman S et al (1991) Increased survival, limb preservation, and prognostic factors for osteosarcoma. Cancer 68:733–737PubMedCrossRefGoogle Scholar
  6. 6.
    Taylor WF, Ivins JC, Unni KK et al (1989) Prognostic variables in osteosarcoma: a multi-institutional study. J Natl Cancer Inst 81:21–30PubMedCrossRefGoogle Scholar
  7. 7.
    Bacci G, Ferrari S, Delepine N et al (1998) Predictive factors of histologic response to primary chemotherapy in osteosarcoma of the extremity: study of 272 patients preoperatively treated with high-dose methotrexate, doxorubicin, and cisplatin. J Clin Oncol 16:658–663PubMedGoogle Scholar
  8. 8.
    Hauben EI, Weeden S, Pringle J et al (2002) Does the histological subtype of high-grade central osteosarcoma influence the response to treatment with chemotherapy and does it affect overall survival? A study on 570 patients of two consecutive trials of the European Osteosarcoma Intergroup. Eur J Cancer 38:1218–1225PubMedCrossRefGoogle Scholar
  9. 9.
    Bacci G, Longhi A, Versari M et al (2006) Prognostic factors for osteosarcoma of the extremity treated with neoadjuvant chemotherapy: 15-year experience in 789 patients treated at a single institution. Cancer 106:1154–1161PubMedCrossRefGoogle Scholar
  10. 10.
    Hauben EI, Bielack S, Grimer R et al (2006) Clinico-histologic parameters of osteosarcoma patients with late relapse. Eur J Cancer 42:460–466PubMedCrossRefGoogle Scholar
  11. 11.
    Hauben EI, Arends J, Vandenbroucke JP et al (2003) Multiple primary malignancies in osteosarcoma patients. Incidence and predictive value of osteosarcoma subtype for cancer syndromes related with osteosarcoma. Eur J Hum Genet 11:611–618PubMedCrossRefGoogle Scholar
  12. 12.
    Svoboda M, Thalhammer T, Aust S et al (2007) Estrogen sulfotransferase (SULT1E1) expression in benign and malignant human bone tumors. J Surg Oncol 95:572–581PubMedCrossRefGoogle Scholar
  13. 13.
    Bilban M, Ghaffari N, Hintermann E et al (2004) Kisspeptin-10, a KiSS1/metastin-derived dekapeptide, is a physiologic invasion inhibitor of primary human trophoblast. J Cell Sci 117:1319–1328PubMedCrossRefGoogle Scholar
  14. 14.
    Bilban M, Heintel D, Scharl T et al (2006) Deregulated expression of fat and muscle genes in B-cell chronic lymphocytic leukemia with high lipoprotein lipase expression. Leukemia 20:1080–1088PubMedCrossRefGoogle Scholar
  15. 15.
    Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Royal Stat Soc 57:289–300Google Scholar
  16. 16.
    Tian L, Greenberg SA, Kong SW et al (2005) Discovering statistically significant pathways in expression profiling studies. Proc Natl Acad Sci USA 102:13544–13549PubMedCrossRefGoogle Scholar
  17. 17.
    Bacci G, Forni C, Ferrari S et al (2003) Neoadjuvant chemotherapy for osteosarcoma of the extremity: intensification of preoperative treatment does not increase the rate of good histologic response to the primary tumor or improve the final outcome. J Pediatr Hematol Oncol 25:845–853PubMedCrossRefGoogle Scholar
  18. 18.
    Roessner A, Voss B, Rauterberg J et al (1983) Biologic characterization of human bone tumors. II. Distribution of different collagen types in osteosarcoma—a combined histologic, immunofluorescence and electron microscopic study. J Cancer Res Clin Oncol 106:234–239PubMedCrossRefGoogle Scholar
  19. 19.
    Kloen P, Gebhardt MC, Perez-Atayde A et al (1997) Expression of transforming growth factor-beta (TGF-beta) isoforms in osteosarcomas: TGF-beta3 is related to disease progression. Cancer 80:2230–2239PubMedCrossRefGoogle Scholar
  20. 20.
    Nikitovic D, Zafiropoulos A, Katonis P et al (2006) Transforming growth factor-beta as a key molecule triggering the expression of versican isoforms v0 and v1, hyaluronan synthase-2 and synthesis of hyaluronan in malignant osteosarcoma cells. IUBMB Life 58:47–53PubMedCrossRefGoogle Scholar
  21. 21.
    Qing J, Maher VM, Tran H et al (1997) Suppression of anchorage-independent growth and matrigel invasion and delayed tumor formation by elevated expression of fibulin-1D in human fibrosarcoma-derived cell lines. Oncogene 15:2159–2168PubMedCrossRefGoogle Scholar
  22. 22.
    Walters DK, Steinmann P, Langsam B et al (2008) Identification of potential chemoresistance genes in osteosarcoma. Anticancer Res 28:673–679PubMedGoogle Scholar
  23. 23.
    Modrowski D, Orosco A, Thévenard J et al (2005) Syndecan-2 overexpression induces osteosarcoma cell apoptosis: implication of syndecan-2 cytoplasmic domain and JNK signaling. Bone 37:180–189PubMedCrossRefGoogle Scholar
  24. 24.
    Orosco A, Fromigué O, Bazille C et al (2007) Syndecan-2 affects the basal and chemotherapy-induced apoptosis in osteosarcoma. Cancer Res 67:3708–3715PubMedCrossRefGoogle Scholar
  25. 25.
    Wilkie AO, Patey SJ, Kan SH et al (2002) FGFs, their receptors, and human limb malformations: clinical and molecular correlations. Am J Med Genet 112:266–278PubMedCrossRefGoogle Scholar
  26. 26.
    Mendoza S, David H, Gaylord GM, Miller CW (2005) Allelic loss at 10q26 in osteosarcoma in the region of the BUB3 and FGFR2 genes. Cancer Genet Cytogenet 158:142–147PubMedCrossRefGoogle Scholar
  27. 27.
    Lorenzi MV, Horii Y, Yamanaka R et al (1996) FRAG1, a gene that potently activates fibroblast growth factor receptor by C-terminal fusion through chromosomal rearrangement. Proc Natl Acad Sci USA 93:8956–8961PubMedCrossRefGoogle Scholar
  28. 28.
    Trieb K, Kotz R (2001) Proteins expressed in osteosarcoma and serum levels as prognostic factors. Int J Biochem Cell Biol 33:11–17PubMedCrossRefGoogle Scholar
  29. 29.
    Mintz MB, Sowers R, Brown KM et al (2005) An expression signature classifies chemotherapy-resistant pediatric osteosarcoma. Cancer Res 65:1748–1754PubMedCrossRefGoogle Scholar
  30. 30.
    Fellenberg J, Dechant MJ, Ewerbeck V, Mau H (2003) Identification of drug-regulated genes in osteosarcoma cells. Int J Cancer 105:636–643PubMedCrossRefGoogle Scholar
  31. 31.
    Han EK, Tahir SK, Cherian SP et al (2000) Modulation of paclitaxel resistance by annexin IV in human cancer cell lines. Br J Cancer 83:83–88PubMedCrossRefGoogle Scholar
  32. 32.
    Cole PD, Kamen BA, Gorlick R et al (2001) Effects of overexpression of gamma-Glutamyl hydrolase on methotrexate metabolism and resistance. Cancer Res 61:4599–4604PubMedGoogle Scholar
  33. 33.
    Tait CR, Jones PF (2004) Angiopoietins in tumours: the angiogenic switch. J Pathol 204:1–10PubMedCrossRefGoogle Scholar
  34. 34.
    Kalinski T, Krueger S, Sel S et al (2006) Differential expression of VEGF-A and angiopoietins in cartilage tumors and regulation by interleukin-1beta. Cancer 106:2028–2038PubMedCrossRefGoogle Scholar
  35. 35.
    Grant DS, Yenisey C, Rose RW et al (2002) Decorin suppresses tumor cell-mediated angiogenesis. Oncogene 21:4765–4777PubMedCrossRefGoogle Scholar
  36. 36.
    Shintani K, Matsumine A, Kusuzaki K et al (2008) Decorin suppresses lung metastases of murine osteosarcoma. Oncol Rep 19:1533–1539PubMedGoogle Scholar
  37. 37.
    Hall CL, Bafico A, Dai J et al (2005) Prostate cancer cells promote osteoblastic bone metastases through Wnts. Cancer Res 65:7554–7560PubMedGoogle Scholar
  38. 38.
    Hoang BH, Kubo T, Healey JH et al (2004) Dickkopf 3 inhibits invasion and motility of Saos-2 osteosarcoma cells by modulating the Wnt-beta-catenin pathway. Cancer Res 64:2734–2739PubMedCrossRefGoogle Scholar
  39. 39.
    Chan KW, Lee PY, Lam AK et al (2006) Clinical relevance of Fas expression in oesophageal squamous cell carcinoma. J Clin Pathol 59:101–104PubMedCrossRefGoogle Scholar
  40. 40.
    Koshkina NV, Khanna C, Mendoza A et al (2007) Fas-negative osteosarcoma tumor cells are selected during metastasis to the lungs: the role of the Fas pathway in the metastatic process of osteosarcoma. Mol Cancer Res 5:991–999PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Bernd Kubista
    • 1
  • Florian Klinglmueller
    • 2
  • Martin Bilban
    • 2
  • Martin Pfeiffer
    • 1
  • Richard Lass
    • 1
  • Alexander Giurea
    • 1
  • Phillipp T. Funovics
    • 1
  • Cyril Toma
    • 1
  • Martin Dominkus
    • 1
  • Rainer Kotz
    • 1
  • Theresia Thalhammer
    • 3
  • Klemens Trieb
    • 4
  • Teresa Zettl
    • 1
  • Christian F. Singer
    • 5
  1. 1.Department of OrthopedicsMedical University of ViennaViennaAustria
  2. 2.Department of Laboratory Medicine and Ludwig Boltzmann Institute for Clinical and Experimental OncologyMedical University of ViennaViennaAustria
  3. 3.Institute of Pathophysiology, Center for Physiology, Pathophysiology and ImmunologyMedical University of ViennaViennaAustria
  4. 4.Department of OrthopedicsKlinikum Kreuzschwestern WelsWelsAustria
  5. 5.Division of Special Gynecology, Department of OB/GYNMedical University of ViennaViennaAustria

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