Abstract
In vitro attachment assays were carried out to assess adhesion between two basement membrane proteins, type IV collagen and laminin, and rat rhabdomyosarcoma (RMS) cell lines with different metastatic potentials. Whereas cells did not adhere to type IV collagen, adhesion to laminin appeared to be very sensitive as maximal adhesion was achieved in dose-response assays with only nanograms of laminin. Adhesion was mediated by interactions between coated laminin and cell surface components, probably receptors, but not endogenous laminin. Laminin-mediated adhesion of RMS cell lines was compared with that of the MCF-7 (human mammary carcinoma) and the L6 (rat myoblast) cell lines. In dose-response assays, RMS cell lines required 10 times less laminin to reach half-maximal attachment rates than MCF-7 and L6 cell lines. Two laminin fragments, P1 and E8, which are structurally and immunologically distinct as shown byα-helix content, SDS-PAGE and monoclonal antibody mapping, supported adhesion by RMS cells and L6 myoblasts, but MCF-7 adhered only to P1. This fragment was 10 times less active than laminin in RMS cell lines. Attachment in dose-response assays and adhesion inhibition studies by antibodies revealed that E8 accounted for the activity of laminin in RMS cell adhesion. Adhesion in the RMS cell lines was dominated by interaction with E8 regardless of metastatic potential.
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Aumailley, M., Nurcombe, V., Edgar, D., Paulsson, M., andTimpl, R., 1987, The cellular interactions of laminin fragments: cell adhesion correlates with two fragment-specific high affinity binding sites.Journal of Biological Chemistry,262, 11532–11538.
Barsky, S. H., Rao, C. N., Hyams, D., andLiotta, L. A., 1984, Characterization of a laminin receptor from human breast carcinoma tissue.Breast Cancer Research and Treatment,4, 181–188.
Charonis, A. S., Tsilibary, E. C., Saku, T., andFurthmayr, H., 1986, Inhibition of laminin self-assembly and interaction with type IV collagen by antibodies to the terminal domain of the long arm.Journal of Cell Biology,103, 1689–1697.
Charpin, C., Lissitzky, J. C., Kopp, F., Pourreau, N., Lavaut, M. N., Martin, P. M., andToga, M., 1986, Immunohistochemical detection of laminin in 92 breast cancers.Human Pathology,17, 355–365.
Couchman, J. R., Höök, M., Rees, D. A., andTimpl, R., 1983, Adhesion, growth, and matrix production by fibroblasts on laminin substrates.Journal of Cell Biology,96, 177–183.
Goodman, S. L., andNewgreen, D., 1985, Do cells show an inverse locomotory response to fibronectin and laminin substrates?EMBO Journal,4, 2769–2771.
Goodman, S. L., Deutzmann, R., andvon der Mark, K., 1987, Two distinct cell-binding domains in laminin can independently promote nonneuronal cell adhesion and spreading.Journal of Cell Biology,105, 589–598.
Graf, J., Iwamoto, Y., Sasaki, M., Martin, G. R., Kleinman, H. K., Robey, F. A., andYamada, Y., 1987, Identification of an amino acid sequence in laminin mediating cell attachment, chemotaxis, and receptor binding.Cell,48, 989–996.
Graf, J., Ogle, R. C., Robey, F. A., Sasaki, M., Martin, G. R., Yamada, Y., andKleinman, H. K., 1987, A pentapeptide from the laminin B1 chain mediates cell adhesion and binds the 67000 laminin receptor.Biochemistry,26, 6896–6900.
Horwitz, A., Duggan, K., Greggs, R., Decker, C., andBuck, C., 1985, The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin.Journal of Cell Biology,101, 2134–2144.
Iwamoto, Y., Robey, F. A., Graf, J., Sasaki, M., Kleinman, H. K., Yamada, Y., andMartin, G. R., 1987, YIGSR, a synthetic laminin pentapeptide, inhibits experimental metastasis formation.Science,238, 1132–1134.
Kleinman, H. K., McGarvey, M. L., Liotta, L. A., Robe, P. G., Tryggvason, K., andMartin, G. R., 1982, Isolation and characterization type IV procollagen, laminin and heparan sulfate proteoglycan from the EHS sarcoma.Biochemistry,21, 6188–6193.
Kramer, R. H., Gonzalez, R., andNicolson, G. L., 1980, Metastatic tumor cells adhere preferentially to the extracellular matrix underlying vascular endothelial cells.International Journal of Cancer,26, 639–645.
Kühl, U., Timpl, R., andvon 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.
Lesot, H., Kühl, U., andvon der Mark, K., 1983, Isolation of a laminin binding protein from muscle cell membranes.EMBO Journal,2, 861–865.
Liotta, L. A., Horan Hand, P., Rao, C. N., Bryant, G., Barsky, S. H., andSchlom, J., 1985, Monoclonal antibodies to the human laminin receptor recognize structurally distinct sites.Experimental Cell Research,156, 117–126.
Lissitzky, J. C., Charpin, C., Bignon, C., Bouzon, M., Kopp, F., Delori, P., andMartin, P. M., 1988, Laminin biosynthesis in the extracellular matrix-producing cell line PFHR9 studied with monoclonal and polyclonal antibodies.Biochemical Journal,250, 843–852.
Mage, M. G., 1980, Preparation of Fab fragments from IgGs of different animal species.Methods in Enzymology, Vol. 70, edited by H. V. Vunakis and J. J. Langone (New York: Academic Press), pp. 142–150.
Malinoff, H. L., McCoy, J. P., Varani, J., andWicha, M. S., 1984, Metastatic potential of murine fibrosarcoma cells is influenced by cell surface laminin.International Journal of Cancer,33, 651–655.
Öcalan, M., Goodman, S. L., Kühl, U., Hauschka, S. D., andvon der Mark, K., 1988, Laminin alters cell shape and stimulates motility and proliferation of murine skeletal myoblasts.Developmental Biology,125, 158–167.
Paulsson, M., Aumailley, M., Deutzmann, R., Timpl, R., Beck, K., andEngel, J., 1987, Laminin-nidogen complex, extraction with chelating agents and structural characterization.European Journal of Biochemistry,166, 11–19.
Poupon, M. F., Becker, M., Verrelle, P., Pauwels, C., Breillout, F., Antoine, E. Lascaux, V., Rolland, Y., Korach, S., Courtois, Y., Moczar, E., Redini, F. Nolibe, D., andMartin, P. M., 1986, The metastatic function of cancer cells as revealed by a rat sarcoma model.Cancer Review,5, 50–82.
Sasaki, M., Kato, S., Kohno, K., Martin, G. R., andYamada, Y., 1987, Sequence of the cDNA encoding the laminin B1 chain reveals a multidomain protein containing cysteine-rich repeats.Proceedings of the National Academy of Sciences of the United States of America,84, 935–939.
Terranova, V. P., Rao, C. N., Kalebic, T., Margulies, I. M., andLiotta, L. A., 1983, Laminin receptor on human breast carcinoma cells.Proceedings of the National Academy of Sciences of the United States of America,80, 444–448.
Timpl, R., Rohde, H., Robey, P. G., Rennard, S. I., Foidart, J. M., andMartin, G. R., 1979, Laminin—a glycoprotein from basement membranes.Journal of Biological Chemistry,254, 9933–9937.
Timpl, R., Johansson, S., van Delden, V., Oberbäumer, I., andHöök, M., 1983, Characterization of protease-resistant fragments of laminin mediating attachment and spreading of rat hepatocytes.Journal of Biological Chemistry,258, 8922–8927.
Vlodavsky, I., andGospodarowicz, D., 1981, Respective roles of laminin and fibronectin in adhesion of human carcinoma and sarcoma cells.Nature (London),289, 304–306.
Wicha, M. S., andHuard, T. K., 1983, Macrophages express cell surface laminin.Experimental Cell Research,143, 475–479.
Yurchenco, P. D., Tsilibary, E. C., Charonis, A. S., andFurthmayr, H., 1985, Laminin polymerizationin vitro, evidence for a two-step assembly with domain specificity.Journal of Biological Chemistry,260, 7636–7644.
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Lissitzky, J.C., Bouzon, M., Loret, E. et al. Laminin-mediated adhesion in metastatic rat rhabdomyosarcoma cell lines involves prominent interactions with the laminin E8 fragment. Clin Exp Metast 7, 469–480 (1989). https://doi.org/10.1007/BF01753667
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DOI: https://doi.org/10.1007/BF01753667