Advertisement

Global Protein-expression Analysis of Bone and Soft Tissue Sarcomas

  • Akira Kawai
  • Tadashi Kondo
  • Yoshiyuki Suehara
  • Kazutaka Kikuta
  • Setsuo Hirohashi
Symposium: Molecular Genetics in Sarcoma

Abstract

Analysis of global protein expression, an approach known as expression proteomics, can offer important clues for understanding tumor biology that cannot be obtained by other approaches (e.g., genome or transcriptome analysis). Using two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry, we performed global protein expression studies of bone and soft tissue sarcomas to develop novel diagnostic and therapeutic biomarkers and allow molecular classification of the tumors. Among 1500 protein variants identified in the two-dimensional gel, 67 proteins correctly distinguished the eight subtypes of 99 histologically classified soft tissue sarcomas. Hierarchical clustering demonstrated leiomyosarcoma and MFH shared a similar protein expression profile, and clear cell sarcoma, synovial sarcoma, and MPNST could be grouped according to their protein expression patterns. Pleomorphic leiomyosarcoma and MFH showed similar tropomyosin isoform expression patterns. Patients with gastrointestinal stromal tumors expressing pfetin protein had better survival than those whose tumors lacked it. We identified 10 protein spots associated with the chemosensitivity of osteosarcoma to preoperative chemotherapy. These 10 spots could be new diagnostic and prognostic markers for osteosarcoma and new therapeutic targets for the disease. Proteomic analysis using 2D-DIGE provides novel information on the biology of bone and soft tissue sarcomas that could be used to diagnosis and treat these tumors.

Level of Evidence: Level II, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence.

Keywords

Osteosarcoma Soft Tissue Sarcoma Protein Spot Synovial Sarcoma Malignant Peripheral Nerve Sheath Tumor 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We thank Robert Nakayama, Eisuke Kobayashi, and Ako Hosono for helpful and stimulating discussions and Misuzu Mori for technical assistance.

References

  1. 1.
    Bacci G, Ferrari S, Delepine N, Bertoni F, Picci P, Mercuri M, Bacchini P, Brach del Prever A, Tienghi A, Camandone A, Campanacci M. 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. 1998;16:658-663.PubMedGoogle Scholar
  2. 2.
    Bacci G, Longhi A, Fagioli F, Briccoli A, Versari M, Picci P. Adjuvant and neoadjuvant chemotherapy for osteosarcoma of the extremities: 27 year experience at Rizzoli Institute, Italy. Eur J Cancer. 2005;41:2836–2845.PubMedCrossRefGoogle Scholar
  3. 3.
    Baldini N, Scotlandi K, Barbanti-Brodano G, Manara MC, Maurici D, Bacci G, Bertoni F, Picci P, Sottili S, Campanacci M, Serra M. Expression of p-glycoprotein in high-grade osteosarcoma in relation to clinical outcome. N Engl J Med. 1995;333:1380–1385.PubMedCrossRefGoogle Scholar
  4. 4.
    Chen G, Gharib TG, Huang CC, Taylor JMG, Misek DE, Kardia SLR, Giordano TJ, Iannettoni MD, Orringer MB, Hanash SM, Beer DG. Discordant protein and mRNA expression in lung adenocarcinomas. Mol Cell Proteomics. 2002;1:304–313.PubMedCrossRefGoogle Scholar
  5. 5.
    Cormier JN, Pollock RE. Soft tissue sarcomas. CA Cancer J Clin. 2004;54:94–109.PubMedCrossRefGoogle Scholar
  6. 6.
    de Alava E. Molecular pathology in sarcomas. Clin Transl Oncol. 2007;9:130–144.PubMedCrossRefGoogle Scholar
  7. 7.
    Fletcher CDM, Gustafson P, Rydholm A, Willen H, Akerman M. Clinicopathologic re-evaluation of 100 malignant fibrous histiocytomas: prognostic relevance of subclassification. J Clin Oncol. 2001;19:3045–3050.PubMedGoogle Scholar
  8. 8.
    Goodman VL, Rock EP, Dagher R, Ramchandani RP, Abraham S, Gobburu JVS, Booth BP, Verbois SL, Morse DE, Liang CY, Chidambaram N, Jiang JX, Tang S, Mahjoob K, Justice R, Pazdur R. Approval summary: sunitinib for the treatment of imatinib refractory or intolerant gastrointestinal stromal tumors and advanced renal cell carcinoma. Clin Cancer Res. 2007;13:1367–1373.PubMedCrossRefGoogle Scholar
  9. 9.
    Grier HE, Krailo MD, Tarbell NJ, Link MP, Fryer CJH, Pritchard DJ, Gebhardt MC, Dickman PS, Perlman EJ, Meyers PA, Donaldson SS, Moore S, Rausen AR, Vietti TJ, Miser JS. Addition of ifosfamide and etoposide to standard chemotherapy for Ewing’s sarcoma and primitive neuroectodermal tumor of bone. N Engl J Med. 2003;348:694–701.PubMedCrossRefGoogle Scholar
  10. 10.
    Grimer RJ. Surgical options for children with osteosarcoma. Lancet Oncol. 2005;6:85–92.PubMedCrossRefGoogle Scholar
  11. 11.
    Guillou L, Coindre JM, Bonichon F, Bui NB, Terrier P, Collin F, Vilain MO, Mandard AM, Doussal VL, Leroux A, Jacquemier J, Duplay H, Sastre-Garau X, Costa J. Comparative study of the National Cancer Institute and French Federation of Cancer Centers Sarcoma Group grading systems in a population of 410 adult patients with soft tissue sarcoma. J Clin Oncol. 1997;15:350–362.PubMedGoogle Scholar
  12. 12.
    Gygi SP, Rochon Y, Franza BR, Aebersold R. Correlation between protein and mRNA abundance in yeast. Mol Cell Biol. 1999;19:1720–1730.PubMedGoogle Scholar
  13. 13.
    Hasegawa T, Hasegawa F, Hirose T, Sano T, Matsuno Y. Expression of smooth muscle markers in so called malignant fibrous histiocytomas. J Clin Pathol. 2003;56:666–671.PubMedCrossRefGoogle Scholar
  14. 14.
    Holt GE, Schwartz HS, Caldwell RL. Proteomic profiling in musculoskeletal oncology by MALDI mass spectrometry. Clin Orthop Relat Res. 2006;450:105–110.PubMedCrossRefGoogle Scholar
  15. 15.
    Ifergan I, Meller I, Issakov J, Assaraf YG. Reduced folate carrier protein expression in osteosarcoma: implications for the prediction of tumor chemosensitivity. Cancer. 2003;98:1958–1966.PubMedCrossRefGoogle Scholar
  16. 16.
    Janknecht R. EWS-ETS oncoproteins: the linchpins of Ewing tumors. Gene. 2005;363:1–14.PubMedCrossRefGoogle Scholar
  17. 17.
    Kakar S, Mihalov M, Chachlani NA, Ghosh L, Johnstone H. Correlation of c-fos, p53, and PCNA expression with treatment outcome in osteosarcoma. J Surg Oncol. 2000;73:125–126.PubMedCrossRefGoogle Scholar
  18. 18.
    Kasper B, Lehnert T, Bernd L, Mechtersheimer G, Goldschmidt H, Ho AD, Egerer G. High-dose chemotherapy with autologous peripheral blood stem cell transplantation for bone and soft-tissue sarcomas. Bone Marrow Transplant. 2004;34:37–41.PubMedCrossRefGoogle Scholar
  19. 19.
    Kawai A, Hosono A, Nakayama R, Matsumine A, Matsumoto S, Ueda T, Tsuchiya H, Beppu Y, Morioka H, Yabe H. Clear cell sarcoma of tendons and aponeuroses: a study of 75 patients. Cancer. 2007;109:109–116.PubMedCrossRefGoogle Scholar
  20. 20.
    Kawai A, Muschler GF, Lane JM, Otis JC, Healey JH. Prosthetic knee replacement after resection of a malignant tumor of the distal part of the femur. J Bone Joint Surg Am. 1998;80:636–647.PubMedCrossRefGoogle Scholar
  21. 21.
    Kondo T, Hirohashi S. Application of highly sensitive fluorescent dyes (CyDye DIGE Fluor saturation dyes) to laser microdissection and two-dimensional difference gel electrophoresis (2D-DIGE) for cancer proteomics. Nat Protoc. 2007;1:2940–2956.CrossRefGoogle Scholar
  22. 22.
    Kondo T, Seike M, Mori Y, Fujii K, Yamada T, Hirohashi S. Application of sensitive fluorescent dyes in linkage of laser microdissection and two-dimensional gel electrophoresis as a cancer proteomic study tool. Proteomics. 2003;3:1758–1766.PubMedCrossRefGoogle Scholar
  23. 23.
    Kusuzaki K, Takeshita H, Murata H, Hirata M, Hashiguchi S, Ashihara T, Hirasawa Y. Relation between cellular doxorubicin binding ability to nuclear DNA and histologic response to preoperative chemotherapy in patients with osteosarcoma. Cancer. 1998;82:2343–2349.PubMedCrossRefGoogle Scholar
  24. 24.
    Ladanyi M. Fusions of the SYT and SSX genes in synovial sarcoma. Oncogene. 2001;5755–5762.Google Scholar
  25. 25.
    Lee YF, John M, Edwards S, Clark J, Flohr P, Maillard K, Edema M, Baker L, Mangham DC, Grimer R, Wooster R, Thomas JM, Fisher C, Judson I, Cooper CS. Molecular classification of synovial sarcomas, leiomyosarcomas and malignant fibrous histiocytomas by gene expression profiling. Br J Cancer. 2003;88:510–515.PubMedCrossRefGoogle Scholar
  26. 26.
    Mariani L, Miceli R, Kattan MW, Brennan MF, Colecchia M, Fiore M, Casali PG, Gronchi A. Validation and adaptation of a nomogram for predicting the survival of patients with extremity soft tissue sarcoma using a three-grade system. Cancer. 2005;103:402–408.PubMedCrossRefGoogle Scholar
  27. 27.
    Marina N, Gebhardt M, Teot L, Gorlick R. Biology and therapeutic advances for pediatric osteosarcoma. The Oncologist. 2004;9:422–441.PubMedCrossRefGoogle Scholar
  28. 28.
    Milano A, Apice G, Ferrari E, Fazioli F, de Rosa V, de Luna AS, Iaffaioli RV, Caponigro F. New emerging drugs in soft tissue sarcoma. Crit Rev Oncol Hematol. 2006;59:74–84.PubMedCrossRefGoogle Scholar
  29. 29.
    Mintz MB, Sowers R, Brown KM, Hilmer SC, Mazza B, Huvos AG, Meyers PA, LaFleur B, McDonough WS, Henry MM, Ramsey KE, Antonescu CR, Chen W, Healey JH, Daluski A, Berens ME, MacDonald TJ, Gorlick R, Stephan DA. An expression signature classifies chemotherapy-resistant pediatric osteosarcoma. Cancer Res. 2005;65:1748–1754.PubMedCrossRefGoogle Scholar
  30. 30.
    Nagayama S, Katagiri T, Tsunoda T, Hosaka T, Nakashima Y, Araki N, Kusuzaki K, Nakayama T, Tsuboyama T, Nakamura T, Imamura M, Nakamura Y, Toguchida J. Genome-wide analysis of gene expression in synovial sarcomas using a cDNA microarray. Cancer Res. 2002;62:5859–5866.PubMedGoogle Scholar
  31. 31.
    Nakayama R, Nemoto T, Takahashi H, Ohta T, Kawai A, Seki K, Yoshida T, Toyama Y, Ichikawa H, Hasegawa T. Gene expression analysis of soft tissue sarcomas: characterization and reclassification of malignant fibrous histiocytoma. Mod Pathol. 2007;20:749–759.PubMedCrossRefGoogle Scholar
  32. 32.
    Nielsen TO, West RB, Linn SC, Alter O, Knowling MA, O’Connell JX, Zhu S, Fero M, Sherlock G, Pollack JR, Brown PO, Botstein D, van de Rijn M. Molecular characterization of soft tissue tumours: a gene expression study. Lancet. 2002;359:1301–1307.PubMedCrossRefGoogle Scholar
  33. 33.
    Oberlin O, Rey A, Desfachelles AS, Philip T, Plantaz D, Schmitt C, Plouvier E, Lejars O, Rubie H, Terrier P, Michon J. Impact of high-dose busulfan plus melphalan as consolidation in metastatic Ewing tumors. J Clin Oncol. 2006;24:3997–4002.PubMedCrossRefGoogle Scholar
  34. 34.
    Ochi K, Daigo Y, Katagiri T, Nagayama S, Tsunoda T, Myoui A, Naka N, Araki N, Kudawara I, Ieguchi M, Toyama Y, Toguchida J, Yoshikawa H, Nakamura Y. Prediction of response to neoadjuvant chemotherapy for osteosarcoma by gene-expression profiles. Int J Oncol. 2004;24:647–655.PubMedGoogle Scholar
  35. 35.
    Resendes BL, Kuo SF, Robertson NG, Giersch ABS, Honrubia D, Ohara O, Adams JC, Morton CC. Isolation from cochlea of a novel human intronless gene with predominant fetal expression. J Assoc Res Otolaryngol. 2004;5:185–202.PubMedGoogle Scholar
  36. 36.
    Rosen G, Caparros B, Huvos AG, Kosloff C, Nirenberg A, Cacavio A, Marcove RC, Lane JM, Mehta B, Urban C. Preoperative chemotherapy for osteogenic sarcoma: selection of postoperative adjuvant chemotherapy based on the response of the primary tumor to preoperative chemotherapy. Cancer. 1982;15:1221–1230.CrossRefGoogle Scholar
  37. 37.
    Suehara Y, Kondo T, Fujii K, Hasegawa T, Kawai A, Seki K, Beppu Y, Nishimura T, Kurosawa H, Hirohashi S. Proteomic signatures corresponding to histological classification and grading of soft-tissue sarcomas. Proteomics. 2006;6:4402–4409.PubMedCrossRefGoogle Scholar
  38. 38.
    Suehara Y, Kondo T, Seki K, Shibata T, Fujii K, Goto M, Hasegawa T, Shimada Y, Sasako M, Shimoda T, Kurosawa H, Beppu Y, Kawai A, Hirohashi S. Pfetin, as a prognostic biomarker of gastrointestinal stromal tumors revealed by proteomics. Clin Cancer Res. 2008;14:1707–1717.PubMedCrossRefGoogle Scholar
  39. 39.
    Tschoep K, Kohlmann A, Schlemmer M, Haferlach T, Issels RD. Gene expression profiling in sarcomas. Crit Rev Oncol Hematol. 2007;63:111–124.PubMedCrossRefGoogle Scholar
  40. 40.
    Tunn PU, Schmidt-Peter P, Pomraenke D, Hohenberger P. Osteosarcoma in children: long-term functional analysis. Clin Orthop Relat Res. 2004;421:212–217.PubMedCrossRefGoogle Scholar
  41. 41.
    Van der Zwan SM, DeMatteo RP. Gastrointestinal stromal tumor: 5 years later. Cancer. 2005;104:1781–1788.PubMedCrossRefGoogle Scholar

Copyright information

© The Association of Bone and Joint Surgeons 2008

Authors and Affiliations

  • Akira Kawai
    • 1
  • Tadashi Kondo
    • 2
  • Yoshiyuki Suehara
    • 1
    • 3
  • Kazutaka Kikuta
    • 1
  • Setsuo Hirohashi
    • 2
  1. 1.Division of Orthopaedic SurgeryNational Cancer Center HospitalTokyoJapan
  2. 2.Proteome Bioinformatics ProjectNational Cancer Center Research InstituteTokyoJapan
  3. 3.Department of Orthopaedic SurgeryJuntendo University School of MedicineTokyoJapan

Personalised recommendations