Three lines of B16 melanoma cells (B16-F1, B16-F10 and B16-BL6) were examined for motility in the micropore filter assay and for synthesis in culture of the basal lamina glycoprotein laminin. All three lines synthesized laminin as judged by the incorporation of [35S]methionine into immunoreactive laminin and secreted (or shed) laminin into the culture medium as indicated by biosynthetic labeling studies and enzyme-linked immunosorbent assays. Immunoreactive laminin was also seen on the surface of the cells as indicated by immunofluorescence staining and by complement-mediated killing. Analysis of [35S]methionine-labeled laminin immunoprecipitates by sodium dodecylsulfatepolyacrylamide gel electrophoresis (SDS-PAGE) both with and without reduction of intersubunit disulfide bonds revealed that all three cell lines produced a similar array of laminin forms, and that the M r=950kD laminin molecule (but not the uncombined subunits) was secreted into the culture medium. Laminin biosynthesis appeared to be limited by the availability of the M r=400kD A subunit as shown by the intracellular accumulation of excess B subunit in the form of uncombined B subunit (M r =200kD) and as a disulfide-linked B dimer (M r=400 kD). The motility of all three cell lines was stimulated four- to five-fold by the addition of either exogenous laminin from the EHS sarcoma or culture medium from the B16 cells containing the secreted laminin. The stimulated motility was inhibited by antilaminin serum. These observations suggest that the laminin synthesized by the B16 melanoma cells themselves may facilitate their motility.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Albrechtsen, R., Nielson, M., Wewer, U., Engvall, E., and Ruoslahti, E., 1981, Basement membrane changes in breast cancer detected by immunochemical staining for laminin. Cancer Research, 41, 5076–5081.
Alitalo, K., Kuismanen, E., Myllyla, R., Kiistala, U., Asko-seljavaara, S., and Vaheri, A., 1982, Extracellular matrix proteins of human epidermal keratinocytes and feeder 3T3 cells. Journal of Cell Biology, 94, 497–505.
Barski, G., and Belehradek, J., jr, 1965, E'tude microcinematographique du mecanisme d'invasion cancereuse en cultures de tissue normal associe aux cellules malignes. Experimental Cell Research, 37, 464–480.
Carr, J., Carr, I., Dreker, B., and Betts, K., 1980, Lymphatic metastasis; invasion of lymphatic vessels and efflux of tumor cells in the afferent popliteal lymph as seen in the Walker rat carcinoma. Journal of Pathology, 132, 287–293.
Carr, I., 1983, Experimental lymphatic metastasis. Journal of Microscopy, 131, 211–220.
Cooper, A. R., Kurkinen, M., Taylor, A., and Hogan, B., 1981, Studies on the biosynthesis of laminin by murine parietal endoderm cells. European Journal of Biochemistry, 119, 189–197.
Fidler, I. J., 1973, Selection of successive tumor lines for metastasis. Nature New Biology, 242, 148–149.
Foidart, J. M., Bere, J. J., Paglia, L., Rennard, S. I., Gulling, M., Martin, G. R., and Katz, S. I., 1980, Distribution and immunoelectron microscopic localization of laminin, a noncollagenous basement membrane glycoprotein. Laboratory Investigation, 42,336–342.
Goldrosen, M. H., and Maslow, D. E., 1985, Relationship between the colonization potential of murine colon adenocarcinoma-38 (MCA-38) cells and in vitro motility. Proceedings of the American Association for Cancer Research, 26, 57.
Haemmerli, G., and Strauli, P., 1978, Motility of L5222 leukemia cells within the mesentery. A study of time-lapse cinematography. Virchows Archiv B. Cell Pathology, 29,167–171.
Hart, I. R., 1979, The selection and characterization of an invasive variant of the B16 melanoma. American Journal of Pathology, 97, 585–600.
Hiserodt, J. C., Laybourn, K. A., and Varani, J., 1985, Expression of a laminin-like substance on the surface of murine natural killer (NK) lymphocytes and its role in NK recognition of tumor target cells. Journal of Immunology, 135, 1484–1487.
Howe, C. C., and Solter, D., 1981, Identification of non-collagenous basement membrane glycopeptides synthesized by mouse parietal endoderm and an endodermal cell line. Developmental Biology, 77, 480–487.
Hujanen, E. S., and Terranova, V. P., 1985, Migration of tumor cells to organ-derived chemoattractants. Cancer Research, 45, 3517–3521.
Kuhl, U., Timpl, R., and Von der Mark, K., 1982, Synthesis of type IV collagen and laminin in cultures of skeletal muscle cells and their assembly on the surface of myotubes. Developmental Biology, 93, 344–354.
Laemmli, U. K., 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.
Mccarthy, J. B., and Furcht, L. T., 1984, Laminin and fibronectin promote the haptotactic migration of B16 melanoma cells in vitro. Journal of Cell Biology, 98, 1474–1480.
Mccoy, J. P., Lloyd, R. V., Wicha, M. S., and Varani, J., Identification of a lamininlike substance on the surface of high-malignant murine fibrosarcoma cells. Journal of Cell Science, 65, 139–151.
Orr, F. W., Varani, J., Delikatny, J., Jain, N., and Ward, P. A., 1981, Comparison of the chemotactic responsiveness of fibrosarcoma subpopulations of differing malignancy. American Journal of Pathology, 102, 160–167.
Peters, B. P., Hartle, R. J., Krzesicki, R. F., Kroll, T. G., Perini, F., Balun, J. E., Goldstein, I. J., and Ruddon, R. W., 1985, The biosynthesis, processing and secretion of laminin by human carcinoma cells. Journal of Biological Chemistry, 260,14732–14743.
Rag, N. C., Barsky, S. H., Terranova, V. P., and Liotta, L. A., 1983, Isolation of a tumor cell laminin receptor. Biochemical and Biophysical Research Communications, 111, 804–808.
Ronnett, G. V., Knutson, V. P., Kohanski, R. A., Simpson, T. L., and Lane, M. D., 1984, Role of glycosylation in the processing of newly translated insulin proreceptor in 3T3-Ll adipocytes. Journal of Biological Chemistry, 259, 4566–4575.
Ruddon, R. W., Hanson, C. A., and Addison, N. J., 1979, Synthesis and processing of human chorionic gonadotropin subunits in cultured choriocarcinoma cells. Proceedings of the National Academy of Sciences, U.S.A., 76, 5143–5147.
Timpl, R., Rohde, H., Robey, P. G., Rennard, S. I., Foidart, J. M., and Martin, G. R., 1979, Laminin-a glycoprotein from basement membranes. Journal of Biological Chemistry, 254, 9933–9937.
Varani, J., Orr, W., and Ward, P. A., 1978, A comparison of the migration of normal and malignant cells in two assay systems. American Journal of Pathology, 90, 159–172.
Varani, J., 1982, Chemotaxis of metastatic tumor cells. Cancer Metastasis Reviews, 1, 17–28.
Varani, J., Orr, W., and Ward, P. A., 1979, Hydrolytic enzyme activities, migratory activity and in vivo growth and metastatic potential of recent tumor isolates. Cancer Research, 39, 2376–2380.
Varani, J., Lovett, E. J., Elgebaly, S., Lundy, J., and Ward, P. A., 1980, In vitro and in vivo adherence of tumor cell variants correlated with tumor formation. American Journal of Pathology, 101, 345–352.
Varani, J., Lovett, E. J., Mccoy, J. P., Shibata, S., Maddox, D. E., Goldstein, I. J., and Wicha, M., 1983, Differential expression of a laminin-like substance by high and low metastatic tumor cells. American Journal of Pathology, 111, 27–34.
Varani, J., Lovett, E. J., Wicha, M., Malinoff, H., and Mccoy, J. P., 1983 a, Cell surface a-D-galactopyranosyl end groups: use as a marker in the isolation of murine tumor cell lines with varying malignant potential. Journal of the National Cancer Institute, 71, 1251–1286.
Varani, J., Fligiel, S. E. G., and Perone, P., 1985, Directional motility in strongly malignant murine tumor cells. International Journal of Cancer, 35, 559–564.
Vesely, P., and Weiss, R. A., 1973, Cell locomotion and contact inhibition of normal and neoplastic rat cells. International Journal of Cancer, 11, 64–76.
Wood, S., 1958, Pathogenesis of metastasis formation observed in vivo in the rabbit ear chamber. Archives of Pathology, 66, 550–568.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fligiel, S.E.G., Laybourn, K.A., Peters, B.P. et al. Laminin production by murine melanoma cells: possible involvement in cell motility. Clin Exp Metast 4, 259–272 (1986). https://doi.org/10.1007/BF00133591
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00133591