The Role of Fas/FasL in the Metastatic Potential of Osteosarcoma and Targeting this Pathway for the Treatment of Osteosarcoma Lung Metastases

Part of the Cancer Treatment and Research book series (CTAR, volume 152)


Pulmonary metastases remain the main cause of death in patients with Osteosarcoma (OS). In order to identify new targets for treatment, our laboratory has focused on understanding the biological properties of the tumor microenvironment that contribute to or interfere with metastasis. Dysfunction of the Fas/FasL signaling pathway has been implicated in tumor development, and progression. Here we describe the status of Fas expression in murine nonmetastatic K7 and metastatic K7M2 cells and human nonmetastatic SAOS and LM2 and metastatic LM6 OS cells. We demonstrated that Fas expression correlates inversely with metastatic potential. Pulmonary metastases from patients were uniformly Fas supporting the importance of Fas expression to the metastatic potential. Since FasL is constitutively expressed in the lung, our data suggests that Fas+ tumor cells undergo apoptosis and are cleared from the lung. By contrast, Fas tumor cells evade this host defense mechanism and form lung metastases. We confirmed these findings by blocking the Fas pathway using Fas Associated Death Domain Dominant-Negative (FDN). Fas+ cells transfected with FDN were not sensitive to FasL, showed delayed clearance and formed lung metastases. Fas+ cells were also able to form lung metastases in FasL-deficient mice. Using our mouse model systems, we demonstrated that aerosol treatment with liposomal 9-Nitrocamptothecin and Gemcitabine (chemotherapeutic agents known to upregulate Fas expression) increased Fas expression and induced tumor regression in wild type mice. Lung metastases in FasL deficient mice did not respond to the treatment.

We conclude that Fas is an early defense mechanism responsible for clearing invading Fas+ tumor cells from the lung. Fas cells or cells with a nonfunctional Fas pathway evade this defense mechanism and form lung metastases. Therapy that induces Fas expression may therefore be effective in patients with established OS lung metastases. Aerosol delivery of these agents is an ideal way to target treatment to the lung.


Lung Metastasis Lung Nodule K7M2 Cell Aerosol Therapy Form Lung Metastasis 
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.



We are very grateful to Joyce Furlough for her clerical assistance. This work was supported in part by NCI grant CA42992 (ESK) and NIH Core grant CA16672.


  1. 1.
    Marina N, et al. Biology and therapeutic advances for pediatric osteosarcoma. Oncologist. 2004;9(4):422-41.CrossRefPubMedGoogle Scholar
  2. 2.
    Meyers PA, et al. Chemotherapy for nonmetastatic osteogenic sarcoma: The Memorial Sloan-Kettering experience. J Clin Oncol. 1992;10(1):5-15.PubMedGoogle Scholar
  3. 3.
    Kager L, et al. Primary metastatic osteosarcoma: Presentation and outcome of patients treated on neoadjuvant Cooperative Osteosarcoma Study Group protocols. J Clin Oncol. 2003;21(10):2011-8.CrossRefPubMedGoogle Scholar
  4. 4.
    Verschraegen CF, et al. Feasibility, phase I, and pharmacological study of aerosolized liposomal 9-nitro-20(S)-camptothecin in patients with advanced malignancies in the lungs. Ann N Y Acad Sci. 2000;922:352-4.CrossRefPubMedGoogle Scholar
  5. 5.
    Khanna C, et al. The membrane-cytoskeleton linker ezrin is necessary for osteosarcoma metastasis. Nat Med. 2004;10(2):182-6.CrossRefPubMedGoogle Scholar
  6. 6.
    Ferguson WS, Goorin AM. Current treatment of osteosarcoma. Cancer Invest. 2001;19(3):292-315.CrossRefPubMedGoogle Scholar
  7. 7.
    Goorin AM, et al. Phase II/III trial of etoposide and high-dose ifosfamide in newly diagnosed metastatic osteosarcoma: A pediatric oncology group trial. J Clin Oncol. 2002;20(2):426-33.CrossRefPubMedGoogle Scholar
  8. 8.
    Goorin AM, et al. Presurgical chemotherapy compared with immediate surgery and adjuvant chemotherapy for nonmetastatic osteosarcoma: Pediatric Oncology Group Study POG-8651. J Clin Oncol. 2003;21(8):1574-80.CrossRefPubMedGoogle Scholar
  9. 9.
    Bruland OS, Pihl A. On the current management of osteosarcoma: A critical evaluation and a proposal for a modified treatment strategy. Eur J Cancer. 1997;33(11):1725-31.CrossRefPubMedGoogle Scholar
  10. 10.
    Owen-Schaub L, et al. Fas and Fas ligand interactions in malignant disease. Int J Oncol. 2000;17(1):5-12.PubMedGoogle Scholar
  11. 11.
    Nagata S. Apoptosis by death factor. Cell. 1997;88(3):355-65.CrossRefPubMedGoogle Scholar
  12. 12.
    Owen-Schaub LB, et al. Fas and Fas ligand interactions suppress melanoma lung metastasis. J Exp Med. 1998;188(9):1717-23.CrossRefPubMedGoogle Scholar
  13. 13.
    Algeciras-Schimnich A, et al. Molecular ordering of the initial signaling events of CD95. Mol Cell Biol. 2002;22(1):207-20.CrossRefPubMedGoogle Scholar
  14. 14.
    Ferguson TA, Griffith TS. A vision of cell death: Insights into immune privilege. Immunol Rev. 1997;156:167-84.CrossRefPubMedGoogle Scholar
  15. 15.
    Lee HO, Ferguson TA. Biology of FasL. Cytokine Growth Factor Rev. 2003;14(3-4):325-35.CrossRefPubMedGoogle Scholar
  16. 16.
    Green DR, Ferguson TA. The role of Fas ligand in immune privilege. Nat Rev Mol Cell Biol. 2001;2(12):917-24.CrossRefPubMedGoogle Scholar
  17. 17.
    Ferguson TA, Green DR. Fas-ligand and immune privilege: The eyes have it. Cell Death Differ. 2001;8(7):771-2.CrossRefPubMedGoogle Scholar
  18. 18.
    Griffith TS, et al. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science. 1995;270(5239):1189-92.CrossRefPubMedGoogle Scholar
  19. 19.
    Griffith TS, et al. CD95-induced apoptosis of lymphocytes in an immune privileged site induces immunological tolerance. Immunity. 1996;5(1):7-16.CrossRefPubMedGoogle Scholar
  20. 20.
    Moller P, et al. Expression of APO-1 (CD95), a member of the NGF/TNF receptor superfamily, in normal and neoplastic colon epithelium. Int J Cancer. 1994;57(3):371-7.CrossRefPubMedGoogle Scholar
  21. 21.
    Hill LL, et al. Fas ligand: A sensor for DNA damage critical in skin cancer etiology. Science. 1999;285(5429):898-900.CrossRefPubMedGoogle Scholar
  22. 22.
    Zornig M, et al. Loss of Fas/Apo-1 receptor accelerates lymphomagenesis in E mu L-MYC transgenic mice but not in animals infected with MoMuLV. Oncogene. 1995;10(12):2397-401.PubMedGoogle Scholar
  23. 23.
    Jia SF, Worth LL, Kleinerman ES. A nude mouse model of human osteosarcoma lung metastases for evaluating new therapeutic strategies. Clin Exp Metastasis. 1999;17(6):501-6.CrossRefPubMedGoogle Scholar
  24. 24.
    Khanna C, et al. Metastasis-associated differences in gene expression in a murine model of osteosarcoma. Cancer Res. 2001;61(9):3750-9.PubMedGoogle Scholar
  25. 25.
    Khanna C, et al. An orthotopic model of murine osteosarcoma with clonally related variants differing in pulmonary metastatic potential. Clin Exp Metastasis. 2000;18(3):261-71.CrossRefPubMedGoogle Scholar
  26. 26.
    Worth LL, et al. Fas expression inversely correlates with metastatic potential in osteosarcoma cells. Oncol Rep. 2002;9(4):823-7.PubMedGoogle Scholar
  27. 27.
    Koshkina NV, Kleinerman ES. Aerosol gemcitabine inhibits the growth of primary osteosarcoma and osteosarcoma lung metastases. Int J Cancer. 2005;116(3):458-63.CrossRefPubMedGoogle Scholar
  28. 28.
    Gordon N, et al. Corruption of the Fas pathway delays the pulmonary clearance of murine osteosarcoma cells, enhances their metastatic potential, and reduces the effect of aerosol gemcitabine. Clin Cancer Res. 2007;13(15 Pt 1):4503-10.CrossRefPubMedGoogle Scholar
  29. 29.
    Gordon N, et al. Fas expression in lung metastasis from osteosarcoma patients. J Pediatr Hematol Oncol. 2005;27(11):611-5.CrossRefPubMedGoogle Scholar
  30. 30.
    Lafleur EA, et al. Increased Fas expression reduces the metastatic potential of human osteosarcoma cells. Clin Cancer Res. 2004;10(23):8114-9.CrossRefPubMedGoogle Scholar
  31. 31.
    Koshkina NV, et al. 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. 2007;5(10):991-9.CrossRefPubMedGoogle Scholar
  32. 32.
    Koshkina NV, et al. 9-Nitrocamptothecin liposome aerosol treatment of melanoma and osteosarcoma lung metastases in mice. Clin Cancer Res. 2000;6(7):2876-80.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Division of PediatricsChildren’s Cancer Hospital, University of Texas M. D. Anderson Cancer CenterHoustonUSA

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