Skip to main content
SpringerLink
Log in

Inhibition of VLA-4 and up-regulation of TIMP-1 expression in B16BL6 melanoma cells transfected with MHC class I genes

  • Published:
Clinical & Experimental Metastasis Aims and scope Submit manuscript

Abstract

The effect of MHC class I gene transfection on the metastatic properties of B16BL6 melanoma cells was investigated. BL6-8 melanoma cells transfected with H-2K or H-2K, but not H-2D or H-2L, genes showed a dramatic reduction in their ability to generate experimental metastases in immunosuppressed CB6F1 mice. This observation suggested that some changes in the metastatic phenotype may have been induced in the H-2K- transfected melanoma cells. Analyses of adhesive and invasive properties of BL6-8 melanoma cells transfected with H-2 class I genes have been performed. We found that the loss of metastatic properties in the H-2K or H-2K gene-transfected melanoma cells was associated with reduced adherence to endothe-lial cells, laminin and collagen IV, decreased ability to form homotypic cell aggregates and with a complete loss of VLA-4 integrin expression. In addition, BL6-8 melanoma cells transfected with H-2K genes demon-strated reduced ability to invade Matrigel that paralleled up -regulation of TIMP-1 expression. Incubation of untransfected BL6-8 clone or B16F1 cells with 5-azacytidine similarly resulted in up-regulation of TIMP-1, suggesting that the changes in methylation of TIMP-1 gene could be responsible for TIMP-1 expression in the H-2K-transfected BL6-8 melanoma cells. Transfection of BL6-8 cells with the H-2D d /L d genes did not affect their adhesive and invasive properties. Previously we reported that reduction in the metastatic properties of the H-2K b transfected cells was associated with alterations in cell surface carbohydrates with appearance of a-galactosyl epitopes and reduction in cell surface sialylation. The present data indicate that, in addition to changes in cell surface carbohydrates, reduction in adhesive properties and up-regulation of TIMP-1 may be responsible for the observed loss of metastatic potential of BL6-8 cells transfected with the H-2K genes. © Rapid Science Ltd.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Gorelik E, Peppoloni S, Overton R and Herberman R, 1985, Increase in H-2 antigen expression and immunogenicity of BL6 melanoma cells treated with N-methyl-N?-nitro-nitrosoguanidine. Cancer Res, 45, 5341-7.

    Google Scholar 

  2. Tanaka K, Gorelik E, Nozumi N and Jay G, 1988, Rejection of BL6 melanoma induced by the expression of a transfected MHC class I gene. Mol Cell Biol, 8, 1857-61.

    Google Scholar 

  3. Itoh T, Storkus W, Gorelik E and Lotze M, 1994, Partial purification of murine tumor-associated peptide epitopes common to histologically distinct tumors, melanoma and sarcoma, that are presented by H-2Kb molecules and recognized by CD8+ tumor infiltrating lymphocytes. J Immunol, 153, 1202-15.

    Google Scholar 

  4. Gorelik E, Jay G, Kwiatkowski B and Herberman R, 1990, Increased sensitivity to MHC-nonrestricted lysis of BL6 melanoma cells by transfection with class I H-2Kb gene. J Immunol, 145, 1621-32.

    Google Scholar 

  5. Kim M, Herberman R and Gorelik E, 1993, Increased sensitivity to TNF-mediated cytotoxicity of BL6 melanoma cells after H-2Kb gene transfection. J Immunol, 151, 3467-77.

    Google Scholar 

  6. Kim M, Duty L, Herberman R and Gorelik E, 1994, Divergent effects of H-2K and H-2D genes on sensitivity of BL6 melanoma cells to NK cells or TNF-mediated cytotoxicity. Cell Immunol, 155, 358-71.

    Google Scholar 

  7. Gorelik E, Kim M, Duty, Henion T and Galili U, 1993, Control of metastatic properties of BL6 melanoma cells by H-2Kb gene: immunological and nonimmunological mechanisms. Clin Exp Metastasis, 11, 439-52.

    Google Scholar 

  8. Gorelik E, Jay G, Kim M, Hearing V, DeLeo A and McCoy P, 1991, Effects of H-2Kb gene on expression of melanoma-associated antigen and lectin-binding sites on BL6 melanoma cells. Cancer Res, 51, 5212-18.

    Google Scholar 

  9. Leong S, Muller J, Yetter R, Gorelik E, Takami T and Hearing V, 1988, Expression and modulation of a retrovirus-associated antigen by murine melanoma cells. Cancer Res, 48, 4954-8.

    Google Scholar 

  10. Li M, Muller J, Xu F, Hearing V and Gorelik E, 1996, Inhibition of melanoma-associated antigen expression and ecotropic retrovirus production in BL6 melanoma cells transfected with MHC class I genes. Cancer Res, 56, 4464-74.

    Google Scholar 

  11. Muller J, Khan A, Li M, Rao V, Hearing V and Gorelik E, 1996, Phenotypic alterations and loss of melanoma-specific endogenous C-type retrovirus production in BL6 melanoma cells transfected with the H-2Kb gene. Melanoma Res, 6, 101-11.

    Google Scholar 

  12. Li M, Muller J, Rao V, Hearing V, Lueders K and Gorelik E, 1996, Loss of intracisternal A-type retroviral particles in BL6 melanoma cells transfected with MHC class I genes. J General Virol, 77, 2757-65.

    Google Scholar 

  13. Prezioso J, Hearing V, Muller J, Urabe K, Wang N and Gorelik E, 1995, Impairment of the melanogenic pathway in BL6 melanoma cells transfected with class I H-2 genes. Melanoma Res, 5, 15-36.

    Google Scholar 

  14. Gorelik E, Duty L, Anaraki F and Galili U, 1995, Alterations of cell surface carbohydrates and inhibition of metastatic properties of murine melanomas by α1,3 galactosyltransferase gene transfection. Cancer Res, 55, 4168-73.

    Google Scholar 

  15. Auerbach R, Alby L, Grieves J et al. 1982, Monoclonal antibody against angiotensin-converting enzyme: its use as a marker for murine, bovine and human endothelial cells. Proc Natl Acad Sci USA, 86, 8227-31.

    Google Scholar 

  16. Giovanni C, Palmieri G, Nicoletti, G, et al. 1991, Immunological and non-immunological influence of H-2Kb gene transfection on the metastatic ability of B16 melanoma cells. Int J Cancer, 48, 270-6.

    Google Scholar 

  17. Sambrook J, Fritsch E and Maiatiatis T, 1989, Molecular Cloning: a laboratory manual. New York: Cold Spring Harbor Laboratory.

    Google Scholar 

  18. Sun Y, Hegamyer G, Kim H, Sithanadandam K, Li H, Watts R and Colburn N, 1995, Molecular cloning of mouse tissue inhibitor of metalloproteinases-3 and its promoter. J Cell Biol, 270, 19312-19.

    Google Scholar 

  19. Rubio M, Nueda A, Vara A and Lopez A-C, 1992, A single mRNA encodes the ?150 and ?80/70 forms of the a subunit of VLA-4. Eur J Immunol, 22, 1099-102.

    Google Scholar 

  20. Edwards D, Rocheleau H, Sharma R, et al. 1992, Involvement of AP1 and PEA3 binding sites in the regulation of murine tissue inhibitor of metalloproteinases-1 (TIMP-1) transcription. Biochem Biophys Acta, 1171, 41-55.

    Google Scholar 

  21. Qian F, Vaux D and Weissman I, 1994, Expression of the integrin α4β1 on melanoma cells can inhibit the invasive stage of metastasis formation. Cell, 77, 335-47.

    Google Scholar 

  22. Heussen C and Dowdle E, 1980, Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Analyt Biochem, 102, 196-202.

    Google Scholar 

  23. Kohn E and Liotta L, 1995, Molecular insights into cancer invasion: strategies for prevention and intervention. Cancer Res, 55, 1856-62.

    Google Scholar 

  24. Matrisian L, 1990, Metalloproteinases and their inhibitors in matrix remodeling. Trends Gen, 6, 121-5.

    Google Scholar 

  25. Hynes R, 1992, Integrins: versatility, modulation and signaling in cell adhesion. Cell, 69, 11-15.

    Google Scholar 

  26. Heino J, 1996, Biology of tumor cell invasion: interplay of cell adhesion and matrix degradation. Int J Cancer, 65, 717-22.

    Google Scholar 

  27. Altevogt P, Hubbe M, Ruppert M, et al. 1995, The α4 integrin chain is a ligand for α4β7 and α4β1. J Exp Med, 182, 34-42.

    Google Scholar 

  28. Khokha R and Denhardt D, 1989, Matrix metalloproteinases and tissue inhibitor of metalloproteinases: a review of their role in tumorigenesis and tissue invasion. Invasion Metastasis, 9, 391-405.

    Google Scholar 

  29. Khokha R, Waterhouse P, Yagel S, Lala P, et al. 1989, Antisense RNA-induced reduction in murine TIMP levels confers oncogenicity on Swiss 3T3 cells. Science, 243, 947-50.

    Google Scholar 

  30. Khokha R, Zimmer M, Graham G, Lala P and Warehouse P, 1992, Suppression of invasion by inducible expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) in B16-F10 melanoma cells. J Natl Cancer Inst, 84, 1017-22.

    Google Scholar 

  31. Koop S, Khokha R, Schmidt, E, et al. 1994, Overexpression of metalloproteinase inhibitor in B16F10 cells does not affect extravasation but reduces tumor growth. Cancer Res, 54, 4791-7.

    Google Scholar 

  32. Laird P and Jaenisch R, 1994, DNA methylation and cancer. Human Mol Genet, 3, 1487-95.

    Google Scholar 

  33. Jones P, 1996, DNA methylation errors and cancer. Cancer Res, 56, 2463-7.

    Google Scholar 

  34. Calarini L, Marozzi A, Byers H, et al. 1992, Expression of a transfected H-2Kb gene in B16 melanoma cells correlates with suppression of liver metastases in triple immunodeficient mice. Cancer Res, 52, 4036-41.

    Google Scholar 

  35. Lauri D, Giovanni C, Biondelli T, et al. 1993, Decreased adhesion to endothelial cells and matrix proteins of H-2Kb gene transfected tumour cells. Br J Cancer, 68, 862-7.

    Google Scholar 

  36. De Giovanni C, Nicoletti G, Sensi M, Santoni A, et al. 1994, H-2Kb and H-2Db gene transfections in B16 melanoma differently affect non-immunological properties relevant to the metastatic process. Involvement of integrin molecules. J Int Cancer, 59, 269-74.

    Google Scholar 

  37. Pargador A, Feldman M and Eisenbach L, 1989, H-2Kb transfection of B16 melanoma cells results in reduced tumorigenicity and metastatic competence. J Immunogenet, 16, 291-303.

    Google Scholar 

  38. Okahara H, Yagita H, Miyake K and Okumura K, 1994, Involvement of VLA-4 and VCAM-1 in TNF??enhancement of experimental metastasis. Cancer Res, 54 3233-6.

    Google Scholar 

  39. Garofalo A, Chirivi R, Foglieni C, et al. 1995, Involvement of the VLA-4 integrin on melanoma in IL-1-augmented experimental metastases. Cancer Res, 55, 414-19.

    Google Scholar 

  40. Vardi I, Gopas J, Rager-Zisman B, Hammerling G, Segal S and Aboud M, 1992, Differential expression of the H-2Dk MHC class I antigen and tissue inhibitor of metalloproteinases in metastatic and nonmetastatic T-10 fibrosarcoma cells. Invasion Metastasis, 12, 301-8.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. University of Pittsburgh Cancer Institute and Departments of Pathology, Pittsburgh, PA, USA

    Fan Xu, Mengfeng Li & Elieser Gorelik

  2. Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Tim Carlos

  3. Ontario Cancer Institute, Toronto, Ontario, Canada

    Otto Sanchez-Sweatman & Rama Khokha

Authors
  1. Fan Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tim Carlos
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mengfeng Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Otto Sanchez-Sweatman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Rama Khokha
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Elieser Gorelik
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xu, F., Carlos, T., Li, M. et al. Inhibition of VLA-4 and up-regulation of TIMP-1 expression in B16BL6 melanoma cells transfected with MHC class I genes. Clin Exp Metastasis 16, 358–370 (1998). https://doi.org/10.1023/A:1006569631330

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006569631330

Search

Navigation