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Expression and role of integrins in adhesion of human colonic carcinoma cells to extracellular matrix components

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Abstract

We have examined integrin expression and function in the human colon carcinoma cell line HT29, and in clonal sublines derived from the HT29 line. These cells express several different integrin subunits including beta 1, alpha 2,3, 6 and alphav, but do not express the classic alpha S/beta 1 fibronectin receptor. Clonal variation in the pattern of integrin expression was quite limited. The profile of integrin expression correlates well with the adhesive behavior of HT29 cells. Thus the cells adhere well to vitronectin, laminin and type IV collagen, but not at all to fibronectin. Adhesion to collagen was completely blocked by an antibeta 1 monoclonal antibody, indicating that beta 1 integrins mediate this process. Adhesion to laminin was strongly blocked by anti-beta 1 monoclonal or anti-beta 6 monoclonal, suggesting that the alpha 6/beta 1 complex functions in attachment to laminin; this was somewhat surprising since immunoprecipitation experiments indicate that most of the alpha 6 subunit seems to be associated with the beta 4 subunit. Despite their strong adherence to laminin, collagen and vitronectin, HT29 cells are not very motile and, in response to gradients of these proteins, do not migrate nearly as well as CHO cells tested under similar conditions. Since HT29 cells can undergo an enterocyte-like differentiation in glucose-free medium, we compared integrin expression in HT29 and its subclones during the process of differentiation. There was no correlation between the state of differentiation, as assessed by expression of brush-border hydrolases, and the level of expression of any of the integrin subunits measured. Thus the pattern of integrin expression in these colonic tumor cells seems to be a characteristic of the cell line, and is not readily modified by changes in cell growth or differentiation.

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References

  1. Akiyama, S. K., Nagata, K., andYamada, K. M., 1990, Cell surface receptors for extracellular matrix.Biochimica et Biophysica Acta,1031, 91–110.

    Google Scholar 

  2. Akiyama, S. K., Yamada, S., Chien, W. T., andYamada, K. M., 1989, Analysis of fibronectin receptor function with monoclonal antibodies.Journal of Cell Biology,109, 863–875.

    Google Scholar 

  3. Albini, A., Iwamoto, Y., Kleinman, H. K., Martin, G. R., Aaronson, S. A., Kozlowski, J. M., andMcEwan, R. N., 1987, A rapid in vitro assay for quantitating the invasive potential of tumor cells.Cancer Research,47, 3239–3245.

    Google Scholar 

  4. Argravess, W. S., Suzuki, S., Arai, H., Thompson, K., Pierschbacher, M. D., andRuoslahti, E., 1987, Amino acid sequence of the human fibronectin receptor.Journal of Cell Biology,105, 1183–1190.

    Google Scholar 

  5. Bodary, S. C., andMcLean, J. W., 1990, The integrin b1 subunit associates with the vitronectin receptor av subunit to form a novel vitronectin receptor in a human embryonic kidney cell line.Journal of Biological Chemistry,265, 5938–5941.

    Google Scholar 

  6. Brown, P. J., andJuliano, R. L., 1987, Association between fibronectin receptor and the substratum: spare receptors for cell adhesion.Experimental Cell Research,171, 376–388.

    Google Scholar 

  7. Brown, P. J., andJuliano, R. L., 1986, Expression and function of a putative cell surface receptor for fibronectin in human and hamster cell lines.Journal of Cell Biology,103, 1595–1603.

    Google Scholar 

  8. Brown, P. J., andJuliano, R. L., 1985, Monoclonal antibodies to a cell surface glycoprotein selectively inhibit fibronectin mediated cell adhesion.Science,228, 1448–1451.

    Google Scholar 

  9. Bruce, W. R., 1987, Recent hypotheses for the origin of colon cancer.Cancer Research,47, 4237–1242.

    Google Scholar 

  10. Buck, C. A., andHorwitz, A. F., 1987, Cell surface receptors for extracellular matrix molecules.Annual Review of Cell Biology,3, 179–205.

    Google Scholar 

  11. Carlow, D. A., Kerbel, R. S., andElliot, B. E., 1989, Failure of expression of class I major histocompatibility antigens to alter tumor immunogenicity of a spontaneous murine carcinoma.Journal of the National Cancer Institute,81, 759–767.

    Google Scholar 

  12. Chanteret, I., Barbat, A., Dussaulx, E., Brattain, M. G., andZweibaum, A., 1988, Epithelial polarity, villin expression, and enterocytic differentiation of cultured human colon carcinoma cells: a survey of twenty cell lines.Cancer Research,48, 1936–1942.

    Google Scholar 

  13. Cheresh, D. A., Smith, J. W., Cooper, H. M., andQuaranta, V., 1989, A novel vitronectin receptor integrin is responsible for distinct adhesive properties of carcinoma cells.Cell,57, 59–69.

    Google Scholar 

  14. Cheung, E., Brown, P. J., andJuliano, R. L., 1987, Altered type I protein kinase in adhesion defective CHO cell variants.Journal of Cellular Physiology,130, 118–124.

    Google Scholar 

  15. Corbi, A. L., Kishimoto, T. K., Miller, L. J., andSpringer, T. A., 1988, Human leukocyte adhesion glycoprotein MAC-1 (complement receptor type 3, CD 11b)α subunit.Journal of Biological Chemistry,263, 12403–12411.

    Google Scholar 

  16. Dahlqvist, A., 1964, Method for assay of intestinal disaccharidases.Analytical Biochemistry,7, 18–25.

    Google Scholar 

  17. Danilov, Y., andJuliano, R. L., 1989, Phorbol ester modulation of integrin mediated cell adhesion: a post-receptor effect.Journal of Cell Biology,108, 1925–33.

    Google Scholar 

  18. Dufour, S., Duband, J.-L., Humphries, M. J., Obara, M., Yamada, K. M., andThiery, J. P., 1988, Attachment, spreading and locomotion of avian neural crest cells are mediated by multiple adhesion sites on fibronectin molecules.EMBO Journal,7, 2661–2671.

    Google Scholar 

  19. Freed, E., Gailit, J., Van der Geer, P., Ruoslahti, E., andHunter, T., 1989, A novel integrinβ subunit is associated with the vitronectin receptor a subunit (av) in a human osteosarcoma cell line and is a substrate for protein kinase C.Embo Journal,8, 2955–2965.

    Google Scholar 

  20. Gehlsen, K. R., Argraves, W. S., Pierschbacher, M. D., andRuoslahti, E., 1988, Inhibition ofin vitro tumor cell invasion by Arg-Gly-Asp-containing synthetic peptides.Journal of Cell Biology,106, 925–930.

    Google Scholar 

  21. Giancotti, F. G., andRuoslahti, E., 1990, Elevated levels of the a5b1 fibronectin receptor suppress the transformed phenotype of Chinese hamster ovary cells.Cell,60, 849–859.

    Google Scholar 

  22. Goren, A., andLevinthal C., 1960, A fine structure genetic and chemical study of the enzyme alkaline phosphatase.Biochimica et Biophysica Acta,38, 470–483.

    Google Scholar 

  23. Hemler, M. E., andJacobson, J. G., 1987, Cell matrix adhesion-related proteins VLA-1 and VLA-2: regulation of expression on T cells.Journal of Immunology,138, 2941–2948.

    Google Scholar 

  24. Huet, C., Sahuquillo-Merino, C., Coudrier, E., andLouvard, D., 1987, Adsorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation.Journal of Cell Biology,105, 345–357.

    Google Scholar 

  25. Humphries, M. J., Yamada, K. M., andOlden, K., 1988, The investigation of the biological effects of anti-cell adhesive synthetic peptides that inhibit experimental metastasis of B16-F10 murine melanoma cells.Journal of Clinical Investigation,81, 782–790.

    Google Scholar 

  26. Hynes, R. O., 1987, Integrins: A family of cell surface receptors.Cell,48, 549–554.

    Google Scholar 

  27. Ignotz, R. A., andMassague, J., 1987, Cell adhesion protein receptors as targets for transforming growth factor-beta action.Cell,51, 189–197.

    Google Scholar 

  28. Irimura, T., Carlson, D. A., Price, J., Yamori, T., Giavazzi, R., Ota, D. M., andCleary, K. R., 1988, Differential expression of a sialoglycoprotein with an approximate molecular weight of 900,000 on metastatic human colon carcinoma cells growing in culture and in tumor tissues.Cancer Research,48, 2353–2360.

    Google Scholar 

  29. Juliano, R. L., 1987, Membrane receptors for extracellular matrix macromolecules: relationship to cell adhesion and tumor metastasis.Biochimica et Biophysica Acta (Reviews on Cancer),907, 261–278.

    Google Scholar 

  30. Kennel, S. J., Foote, L. J., Falcioni, R., Sonnenberg, A., Sttinger, C. D., Crouse, C., andHemler, M. E., 1989, Analysis of the tumor associated antigen TSP-180; identity with alpha 6/beta 4 in the integrin superfamily.Journal of Biological Chemistry,264, 15515–15521.

    Google Scholar 

  31. Kirchofer, D., Languino, L. R., Duoslahti, E., andDierschbacher, M. D., 1990, Alpha 2/beta 1 integrins from different cell types show different binding specificities.Journal of Biological Chemistry,265, 615–618.

    Google Scholar 

  32. Kishimoto, T. K., Hollander, N., Roberts, T. M., Anderson, D. C., andSpringer, T. A., 1987, Heterogeneous mutations in the beta subunit common to the LFA-1, MAC-1 and p150/95 glycoproteins cause leukocyte adhesion deficiency.Cell,50, 193–202.

    Google Scholar 

  33. Kramer, R. H., McDonald, K. A., andVu, M. P., 1989, Human melanoma cells express a novel integrin receptor for laminin.Journal of Biological Chemistry,264, 15642–15649.

    Google Scholar 

  34. Liotta, L. A., 1986, Tumor invasion and metastasis-role of the extracellular matrix.Cancer Research,46, 1–7.

    Google Scholar 

  35. Lotz, M. M., Korzelius, C. A., andMercurio, A., 1990, Human colon carcinoma cells use multiple receptors to adhere to laminin.Cell Regulation,1, 249–257.

    Google Scholar 

  36. McCarthy, J. B., Basara, M. L., Palm, S. L., Sas, D. F., andFurcht, L. T., 1985, The role of cell adhesion proteins-laminin and fibronectin-in the movement of malignant and metastatic cells.Cancer and Metastasis Review,4, 125–152.

    Google Scholar 

  37. Nicolson, G. L., 1988, Cancer metastasis; tumor cell and host organ properties in metastasis to specific secondary sites.Biochimica et Biophysica Acta,948, 175–224.

    Google Scholar 

  38. Obara, M., Kang, M. S., andYamada, K. M., 1988, Site-directed mutagenesis of the cell-binding domain of human fibronectin: separable, synergistic sites mediate adhesive function.Cell,53, 649–657.

    Google Scholar 

  39. Parise, L.V.,Criss, A. B.,Nannizzi, L.,Wardell, M. R., 1990, Phosphorylation of platelet glycoprotein IIb/IIIa.Blood. (In press).

  40. Phillips, D. R., Charo, I. F., Parise, L. V., andFitzgerald, L. A., 1988, The platelet membrane glycoprotein IIb-IIIa complex.Blood,71, 831–843.

    Google Scholar 

  41. Pierschbacher, M. B., Hayman, E. C., andRuoslahti, E., 1983, Synthetic peptide with cell attachment activity of fibronectin.Proceedings of the National Academy of Science, USA. 80, 1224–1227.

    Google Scholar 

  42. Pierschbacher, M. D., andRuoslahti, E., 1987, Influence of stereochemistry of the sequence Arg-Gly-Asp-Xaa on binding specificity in cell adhesion.Journal of Biological Chemistry,262, 17294–17298.

    Google Scholar 

  43. Plantefaber, L. C., andHynes, R. O., 1989, Changes in integrin receptors on oncogenically transformed cells.Cell,56, 281–290.

    Google Scholar 

  44. Poste, G., andFidler, I. J., 1980, The pathogenesis of cancer metastasis.Nature,283, 139–146.

    Google Scholar 

  45. Pytela, R., Pierschbacher, M. D., andRuoslahti, E., 1985, Identification and isolation of a 140kd cell surface glycoprotein with properties expected of a fibronectin receptor.Cell,40, 191–198.

    Google Scholar 

  46. Ramos, D. M., Berston, E. D., andKramer, R. H., 1990, Analysis of integrin receptors for laminin and type IV collagen on metastatic B16 melanoma cells.Cancer Research,50, 728–734.

    Google Scholar 

  47. Roosien, F. F., de Rifk, D., Bikker, A., andRoos, E., 1989, Involvement of LFA-1 in lymphoma invasion and metastasis demonstrated with LFA-1 deficient mutants.Journal of Cell Biology,108, 1979–1985.

    Google Scholar 

  48. Schreiner, C. L., Bauer, J. S., Danilov, Y. N., Hussein, S., Sczekan, M. M., andJuliano, R. L., 1989, Isolation and characterization of CHO cell variants deficient in the expression of fibronectin receptor.Journal of Cell Biology,109, 3157–3167.

    Google Scholar 

  49. Schwarz, M. A., andJuliano, R. L., 1984, Surface activation of the cell adhesion fragment of fibronectin.Experimental Cell Research,153, 550–555.

    Google Scholar 

  50. Solowska, J., Guan, J.-L., Marcantonio, E. E., Trevithick, J. E., Buck, C. A., andHynes, R. O., 1989, Expression of normal and mutant avian integrin subunits in rodent cells.Journal of Cell Biology,109, 853–861.

    Google Scholar 

  51. Sonnenberg, A., Modderman, P. W., andHogervorst, F., 1988, The laminin receptor on platelets is the integrin VLA-6Nature,336, 487–489.

    Google Scholar 

  52. Soybel, D. I., Bliss, D. P., Jr, 1987, Colon and rectal carcinoma.Current Problems in Cancer,11, 257–356.

    Google Scholar 

  53. Suzuki, S., Argraves, W. S., Pytela, R., Arai, H., Krusius, T., Pierschbacher, M. D., andRuoslahti, E., 1986, cDNA and amino acid sequences of the cell adhesion protein receptor recognizing vitronectin reveal a transmembrane domain and homologies with other adhesion protein receptors.Proceedings of the National Academy of Sciences, USA,83, 8614–8618.

    Google Scholar 

  54. Takeda, Y., Strominger, J. L., andHemler, M. E., 1987, The very late antigen family of heterodimers is part of a superfamily of molecules involved in adhesion and embryogenesis.Proceedings of the National Academy of Sciences, USA,84, 3239–3243.

    Google Scholar 

  55. Tamkun, J. W., DeSimone, D. W., Fonda, D., Pateel, R. S., Buck, C. S., Horwitz, A. F., andHynes, R. O., 1986, Structure of integrin: a glycoprotein involved in transmembrane linkage between fibronectin and actin.Cell,56, 271–282.

    Google Scholar 

  56. Wayner, E. A., andCarter, W. G., 1987, Identification of multiple cell adhesion receptors for collagen and fibronectin in human fibrosarcoma cells possessing unique alpha and beta subunits.Journal of Cell Biology,105, 1873–1884.

    Google Scholar 

  57. Wayner, E. A., Carter, W. G., Piotrowicz, R. S., andKunicki, T. J., 1988, The function of multiple extracellular matrix receptors in mediating cell adhesion to extracellular matrix: Preparation of monoclonal antibodies to the fibronectin receptor that specifically inhibit cell adhesion to fibronectin and react with platelet glycoproteins Ic-IIa.Journal of Cell Biology,107, 1881–1891.

    Google Scholar 

  58. Werb, Z., Tremble, P. M., Behrendtsen, O., Crowley, E., andDamsky, C. H., 1989, Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression.Journal of Cell Biology,109, 877–889.

    Google Scholar 

  59. Zetter, B., 1990, The cellular basis of site specific tumor metastasis.New England Journal of Medicine,322, 605–612.

    Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, University of North Carolina, 27599, Chapel Hill, North Carolina, USA

    C. Schreiner, J. Bauer, M. Margolis & R. L. Juliano

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  1. C. Schreiner
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  3. M. Margolis
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  4. R. L. Juliano
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Schreiner, C., Bauer, J., Margolis, M. et al. Expression and role of integrins in adhesion of human colonic carcinoma cells to extracellular matrix components. Clin Exp Metast 9, 163–178 (1991). https://doi.org/10.1007/BF01756387

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  • DOI: https://doi.org/10.1007/BF01756387

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