Skip to main content

Advertisement

Log in

Use of NeoR B16F1 murine melanoma cells to assess clonality of experimental metastases in the immune-deficient chick embryo

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

Abstract

Recent work on molecular and genetic aspects of metastasis has emphasized the need for assays in immune-deficient animal hosts. The commonly used assays in athymic nude mice may not always be appropriate, and assays in other hosts are required. We have developed a metastasis assay in the naturally immune-deficient chicken embryo. As part of our characterization of this assay we have examined the clonality of individual experimental (i.v.-derived) metastases in this host. For these studies we developed a cell line, B16-Neo, from parental B16F1 murine melanoma cells. B16-Neo cells carry a stable drug-resistance marker, the bacterialneo gene, which confers resistance to the drug G418, but are unaltered in experimental metastatic properties in the chick embryo relative to parental B16F1 cells. We observe that the majority of individual liver tumors that arise following i.v. injection of mixtures of these cells contain cells of a single marker phenotype and are likely to be clonal in origin. These results are similar to those obtained by others for metastases in immune-competent mice, suggesting similar mechanisms of metastasis formation in these two systems. In both hosts it should be noted, however, that a small but significant proportion of metastases appearnot to be clonal in origin.

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

Access this article

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Bondy, G. P., Wilson, S., andChambers, A. F., 1985, Experimental metastatic ability of H-ras-transformed NIH3T3 cells.Cancer Research,45, 6005–6009.

    PubMed  Google Scholar 

  2. Chambers, A. F., 1986, Use of the chick embryo in studying the molecular genetics of metastasis.Biochemistry and Molecular Genetics of Cancer Metastasis, edited by K. Lapis, L. A. Liotta and A. S. Rabson, pp. 11–23.

  3. Chambers, A. F., Harris, J. F., Ling, V., andHill, R. P., 1984, Rapid phenotype variation in cells derived from lung metastases of KHT fibrosarcoma.Invasion and Metastasis,4, 225–237.

    Google Scholar 

  4. Chambers, A. F., andLing, V., 1984, Selection for experimental metastatic ability of heterologous tumor cells in the chick embryo after DNA-mediated transfer.Cancer Research,44, 3970–3975.

    PubMed  Google Scholar 

  5. Chambers, A. F., Shafir, R., andLing, V., 1982, A model system for studying metastasis using the embryonic chick.Cancer Research,42, 4018–4025.

    PubMed  Google Scholar 

  6. Chambers, A. F., andWilson, S., 1985, Cells transformed with ats viralsrc mutant are temperature sensitive forin vivo growth.Molecular and Cellular Biology,5, 728–733.

    PubMed  Google Scholar 

  7. Dick, J. E., Magli, M. C., Huszar, D., Phillips, R. A., andBernstein, A., 1985, Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/Wv mice.Cell,42, 71–79.

    PubMed  Google Scholar 

  8. Fidler, I. J., 1973, Selection of successive tumor lines for metastasis.Nature New Biology,242, 148–149.

    PubMed  Google Scholar 

  9. Fidler, I. J., 1975, Biological behavior of malignant melanoma cells correlated to their survivalin vivo.Cancer Research,35, 218–224.

    PubMed  Google Scholar 

  10. Fidler, I. J., 1985, Macrophages and metastasis, a biological approach to cancer therapy.Cancer Research,45, 4714–4726.

    PubMed  Google Scholar 

  11. Fidler, I. J., 1986, Rationale and methods for the use of nude mice to study the biology and therapy of human cancer metastasis.Cancer Metastasis Reviews,5, 29–49.

    PubMed  Google Scholar 

  12. Fidler, I. J., andHart, I. R., 1982, Biological diversity in metastatic neoplasms: origins and implications.Science,217, 998–1003.

    PubMed  Google Scholar 

  13. Fidler, I. J., andNicolson, G. L., 1976, Organ selectivity for implantation survival and growth of B16 melanoma variant cells lines.Journal of the National Cancer Institute,57, 1199–1201.

    PubMed  Google Scholar 

  14. Fidler, I. J., andTalmadge, J. E., 1986, Evidence that intravenously derived murine pulmonary melanoma metastases can originate from the expansion of a single cell.Cancer Research,46, 5167–5171.

    Google Scholar 

  15. Frost, P., andKerbel, R. S., 1983, Immunology of metastasis. Can the immune system cope with disseminated tumor?Cancer Metastasis Reviews,2, 239–256.

    PubMed  Google Scholar 

  16. Hanna, N., 1982, Role of natural killer cells in control of cancer metastasis.Cancer Metastasis Reviews,1, 45–64.

    PubMed  Google Scholar 

  17. Harris, J. F., Chambers, A. F., Hill, R. P., andLing, V., 1982, Metastatic variants are generated spontaneously at a high rate in mouse KHT tumor.Proceedings of the National Academy of Sciences, U.S.A.,79, 5547–5551.

    Google Scholar 

  18. Heppner, G. H., andMiller, B. E., 1983, Tumor heterogeneity: biological implications and therapeutic consequences.Cancer Metastasis Reviews,2, 5–23.

    PubMed  Google Scholar 

  19. Hill, R. P., Chambers, A. F., Ling, V., andHarris, J. F., 1984, Dynamic heterogeneity: rapid generation of metastatic variants in mouse B16 melanoma cells.Science,224, 998–1001.

    PubMed  Google Scholar 

  20. Joyner, A. L., andBernstein, A., 1983, Retrovirus transduction: generation of infectious retroviruses expressing dominant and selectable genes is associated within vivo recombination and deletion events.Molecular and Cellular Biology,3, 2180–2190.

    PubMed  Google Scholar 

  21. Nicolson, G. L., 1987, Tumor cell instability, diversification, and progression to the metastatic phenotype: from oncogene to oncofetal expression.Cancer Research,47, 1473–1487.

    PubMed  Google Scholar 

  22. Poste, G., Doll, J., Brown, A. E., Tzeng, J., andZeidman, I., 1982, Comparison of the metastatic properties of B16 melanoma clones isolated from cultured cell lines, subcutaneous tumors, and individual lung metastases.Cancer Research,42, 2770–2778.

    PubMed  Google Scholar 

  23. Poste, G., Tzeng, J., Doll, J., Greig, R., Rieman, D., andZeidman, I., 1982, Evolution of tumor cell heterogeneity during progressive growth of individual lung metastases.Proceedings of the National Academy of Sciences, U.S.A.,79, 6574–6578.

    Google Scholar 

  24. Salisbury, A. J., 1975, The significance of the circulating cancer cell.Cancer Treatment Reviews,2, 55–75.

    PubMed  Google Scholar 

  25. Southern, P. J., andBerg, P., 1982, Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.Journal of Molecular and Applied Genetics,1, 327–341.

    PubMed  Google Scholar 

  26. Talmadge, J. E., Wolman, S. R., andFidler, I. J., 1982, Evidence for the clonal origin of spontaneous metastases.Science,217, 361–363.

    PubMed  Google Scholar 

  27. Talmadge, J. E., andZbar, B., 1987, Clonality of pulmonary metastases from the bladder 6 subline of the B16 melanoma studied by Southern hybridization.Journal of the National Cancer Institute,78, 315–320.

    PubMed  Google Scholar 

  28. Thorgeirsson, U. P., Turpeenniemi-Hujanen, T., Williams, J. E., Westin, E. H., Heilman, C. A., Talmadge, J. E., andLiotta, L. A., 1985, NIH/3T3 cells transfected with human tumor DNA containing activatedras oncogenes express the metastatic phenotype in nude mice.Molecular and Cellular Biology,5, 259–262.

    PubMed  Google Scholar 

  29. Trimble, W. S., Johnson, P. W., Hozumi, N., andRoder, J. C., 1986, Inducible cellular transformation by a metallothionein-ras hybrid oncogene leads to natural killer cell susceptibility.Nature,321, 782–784.

    PubMed  Google Scholar 

  30. Weiss, L., 1980, Metastasis: differences between cancer cells in primary and secondary tumors.Pathobiology Annual,10, 51–81.

    PubMed  Google Scholar 

  31. Weiss, L., andGlaves, D., 1983, Cancer cell damage at the vascular endothelium.Annals of the New York Academy of Sciences,416, 681–692.

    PubMed  Google Scholar 

  32. Weiss, L., Dimitrov, D. S., andAngelova, M., 1985, The hemodynamic destruction of intravascular cancer cells in relation to myocardial metastasis.Proceedings of the National Academy of Sciences, U.S.A.,82, 5737–5741.

    Google Scholar 

  33. Welch, D. R., Krizman, D. B., andNicolson, G. L., 1984, Multiple phenotypic divergence of mammary adenocarcinoma cell clones. I.In vitro andin vivo properties.Clinical and Experimental Metastasis,2, 333–355.

    PubMed  Google Scholar 

  34. Woodruff, M. F. A., 1983, Cellular heterogeneity in tumours.British Journal of Cancer,47, 589–594.

    PubMed  Google Scholar 

  35. Young, S. D., andHill, R. P., 1986, Dynamic heterogeneity: isolation of murine tumor cell populations enriched for metastatic variants and quantification of the unstable expression of the phenotype.Clinical and Experimental Metastasis,4, 153–176.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chambers, A.F., Wilson, S. Use of NeoR B16F1 murine melanoma cells to assess clonality of experimental metastases in the immune-deficient chick embryo. Clin Exp Metast 6, 171–182 (1988). https://doi.org/10.1007/BF01784847

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01784847

Keywords

Navigation