Summary
Integrin-mediated tumour cell adhesion to extracellular matrix (ECM) components is an important step in the development of metastatic lesions. Thus, integrin expression and integrin-mediated adhesion of colon carcinoma cells to various ECM components was examined. Poorly (HT-29P) and highly (HT-29LMM) liver-metastatic colon carcinoma cells were used to study the rates of adhesion to collagen I (C I), collagen IV (C IV), laminin (LN), fibronectin (FN), or vitronectin (VN) in a static adhesion assay (10–120 min). Cells were untreated or treated with oligopeptides (RGD, GRGDS, YIGSR, RGES), anti-integrin antibodies, or colchicine, nocodazole, cycloheximide, acrylamide or cytochalasin D (to disrupt cytoskeletal structures). Both cell lines expressed similar patterns of integrin expression (α2, α3, α6, αv, β1, β4 and β5) by immunocytochemistry and immunoprecipitation. HT-29LMM cells showed significantly higher rates of adhesion to LN (P < 0.001) and FN (P < 0.001), but significantly poorer rates of adhesion to C I (P < 0.05) and C IV (P < 0.001) than HT-29P cells, respectively, adhesion to VN was insignificant. RGD and GRGDS inhibited HT-29LMM cell adhesion to FN only. Pretreatment with anti-β1 or anti-α2 integrin subunits suppressed adhesion to C I and C IV, and adhesion to LN was inhibited with anti-β1 or anti-α6 integrin. Anti-β1 or anti-αv blocked adhesion to FN. Pretreatment of cells with cytochalasin D, cycloheximide or acrylamide inhibited adhesive interactions of both cell lines to the ECM components. In contrast, colchicine and nocodazole had no effect. The results demonstrate that adhesion of HT-29 cells to ECM is mediated, in part, by different integrins, depending on the substrate. Poorly and highly metastatic HT-29 cells possessed different patterns of adhesion to the various ECM substrates, but these differences were not due to different expression of integrin subunits. The results also suggested that the initial adhesion of poorly or highly metastatic HT-29 cells to ECM components requires, in part, the presence of native action and intermediate filaments, but not of microtubules. Thus the adhesion of tumour cells to ECM components may be dependent on signal transduction and assembly of microfilaments.
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References
Agrez, M. V. & Bates, R. C. (1994). Colorectal cancer and the integrin family of cell adhesion receptors: Current status and future directions. Eur J Cancer 14: 2166–2170.
Akiyama, S. K., Yamada, S. S., Yamada, K. M. & Laflamme, S. E. (1994). Transmembrane signal transduction by integrin cytoplasmic domains expressed in single-subunit chimeras. J Biol Chem 269: 15961–15964.
Ben-Ze’ev, A. (1997). Cytoskeletal and adhesion proteins as tumor suppressors. Curr Opin Cell Biol 9: 99–108.
Ben-Ze’ev, A. & Raz, A. (1985). Relationship between the organization and synthesis of vimentin and the metastatic capability of B16 melanoma cells. Cancer Res 45: 2632–2641.
Botteri, F. M., Ballmer-Hofer, K., Rajput, B. & Nagamine, Y. (1990). Disruption of cytoskeletal structures results in the induction of the urokinase-type plasminogen activator gene expression. J Biol Chem 265: 13327–13334.
Boyd, D., Florent, G., Childress-Fields, K. & Brattain, M. G. (1988). Alteration in the behavior of a colon carcinoma cell line by extracellular matrix components. Cancer Lett 41: 81–90.
Burtin, P., Chavanel, G. & Foidart, J. M. (1983). Immunofluorescence study of the antigens of the basement membrane and the peritumoral stroma in human colonic adenocarcinomas. Ann NY Acad Sci, 229–236.
Chen, Y. P., O’Toole, T. E., Shipley, T., Forsyth, J., Laflamme, S. E., Yamada, K. M., Shattil, S. J. & Ginsberg, M. H. (1994). ‘Inside-out’ signal transduction inhibited by isolated integrin cytoplasmic domains. J Biol Chem 269: 18307–18310.
Chopra, H., Hatfield, J. S., Chang, Y. S., Grossi, I. M., Fitzgerald, L. A., O’Gara, C. Y., Marnett, L. J., Diglio, C. A., Taylor, J. D. & Honn, K. V. (1988). Role of tumor cytoskeleton and membrane glycoprotein IRGpIIb/IIIa in platelet adhesion to tumor cell membrane and tumor cell-induced platelet aggregation. Cancer Res 48: 3787–3800.
Chopra, H., Fligiel, S. E., Hatfield, J. S., Nelson, K. K., Diglio, C. A., Taylor, J. D. & Honn, K. V. (1990). An in vivo study of the role of the tumor cell cytoskeleton in tumor cell-platelet-endothelial cell interactions. Cancer Res 50: 7686–7696.
Chopra, H., Timar, J., Chen, Y. Q., Rong, X. H., Grossi, I. M., Fitzgerald, L. A., Taylor, J. D. & Honn, K. V. (1991). The lipoxygenase metabolite 12(S)-HETE induces a cytoskeleton-dependent increase in surface expression of integrin αIIbβ3 on melanoma cells. Int J Cancer 49: 774–786.
Danecker, G. W., Piazza, A. J., Stelle, G. D. & Mercurio, A. M. (1989). Relationship between extracellular matrix interactions and degree of differentiation in human colon carcinoma cell lines. Cancer Res 49: 681–686.
Doerr, R., Zvibel, I., Chiuten, D., D’Olimpio, J. & Reid, L. M. (1989). Clonal growth of tumors on tissue-specific biomatrices and correlation with organ site specificity of metastasis. Cancer Res 49: 384–339.
Gong, J., Wang, D. H., Sun, L. Z., Zborowska, E., Willson, J. K. V. & Brattain, M. G. (1997). Role of α5β1 integrin in determining malignant properties of colon carcinoma cells. Cell Growth Diff 8: 83–90.
Grigioni, W. F., Biagini, G., Errico, A. D., Milani, M., Villanacci, V., Garbisa, S., Mattioli, S., Gozzetti, G. & Mancini, A. M. (1986). Behaviour of basement membrane antigens in gastric and colorectal cancer: Immunohistochemical study. Acta Pathol Jpn 36: 173–184.
Guan, J. L., Trevithick, J. E. & Hynes, R. O. (1991). Fibronectin/integrin interaction induces tyrosine phosphorylation of a 120 kDa protein. Cell Regul 2: 951–964.
Gumbiner, B. M. (1996). Cell adhesion: The molecular basis of tissue architecture and morphogenesis. Cell 84: 345–357.
Gurland, G. & Gundersen, G. G. (1995). Stable, detyrosinated microtubules function to localize vimentin intermediate filaments in fibroblasts. J Cell Biol 131: 1275–1290.
Haier, J., Nasralla, M., Buhr, H. J. & Nicolson, G. L. (1998). Differential integrin-associated adhesion to extracellular matrix in colon cancer cells with low metastatic potential or highly metastatic potential to the liver. Langenbecks Arch Chir Suppl, 307–313.
Hendrix, M. J. C., Seftor, E. A., Chu, Y. W., Trevor, K. T. & Seftor, R. E. B. (1996). Role of intermediate filaments in migration, invasion and metastasis. Cancer Metast Rev 15: 507–525.
Hendrix, M. J., Seftor, E. A., Seftor, R. E. & Trevor, K. T. (1997). Experimental co-expression of vimentin and keratin intermediate filaments in human breast cancer cells results in phenotypic interconversion and increased invasive behavior. Am J Pathol 150: 483–495.
Herzberg, F., Schöning, M., Schirner, M., Topp, M., Thiel, E. & Kreuser, E. D. (1996). IL-4 and TNF-α induce changes in integrin expression and adhesive properties and decrease the lung-colonizing potential of HT-29 colon carcinoma cells. Clin Exp Metastasis 14: 165–175.
Horwitz, A., Duggan, K., Buck, C., Beckerle, M. C. & Burrigde, K. (1986). Interaction of plasma membrane fibronectin receptor with talin: a transmembrane linkage. Nature 320: 531–533.
Inagaki, N., Goto, H., Ogawara, M., Nishi, Y., Ando, S. & Inagaki, M. (1997). Spatial patterns of Ca2+ signals define intracellular distribution of a signaling by Ca2+/calmodulin-dependent protein kinase II. J Biol Chem 272: 25195–25199.
Inufusa, H., Nakamura, M., Adachi, T., Nakatani, Y., Shindo, K., Yasutomi, M. & Matsuura, H. (1995). Localization of oncofetal and normal fibronectin in colorectal cancer. Cancer 75: 2802–2808.
Jewell, K., Kapron-Bras, C., Jeevaratnam, P. & Dedhar, S. (1995). Stimulation of tyrosine phosphorylation of distinct proteins in response to antibody-mediated ligation and clustering of α3 and α6 integrins. J Cell Sci 108: 1165–1174.
Kemperman, H., Wijnands, Y. M. & Roos, E. (1997). Alpha V integrins on HT-29 colon carcinoma cells: adhesion to fibronectin is mediated solely by small amounts of alphaVbeta6, and alphaVbeta5 is codistributed with actin fibers. Exp Cell Res 234: 156–164.
Kim, W. H., Jun, S. H., Kibbey, M. C., Thompson, E. W. & Kleinman, H. K. (1994). Expression of beta 1 integrin in laminin-adhesion-selected human colon cancer cell lines of varying tumorigenicity. Invasion Metastasis 14: 147–155.
Kornberg, L. J., Earp, H. S., Turner, C. E., Prockop, C. & Juliano, R. L. (1991). Signal transduction by integrins: Increased protein tyrosine phosphorylation caused by clustering of β1 integrins. Proc Natl Acad Sci USA 88: 8392–8396.
Kreis, T. & Vale, R. (1993). Guidebook of the cytoskeletal and motor proteins. Oxford University Press, Oxford
Lamb, N. J., Fernandez, A., Feramisco, J. R. & Welch, W. J. (1989). Modulation of vimentin containing intermediate filament distribution and phosphorylation in living fibroblasts by the cAMP-dependent protein kinase. J Cell Biol 108: 2409–2422.
Lehmann, M., Rabenandrasana, C., Tamura, R., Lissitzky, J. C., Quaranta, J. P. & Marvaldi, J. (1994). A monoclonal antibody inhibits adhesion to fibronectin and vitronectin of a colon carcinoma cell line and recognizes the integrins αvβ3, αvβ5 and αvβ6. Cancer Res 54: 2102–2107.
Lindmark, G., Gerdin, B., Pahlman, L., Glimelius, B., Gehlsen, K. & Rubin, K. (1993). Interconnection of integrins alpha 2 and alpha 3 and structure of the basal membrane in colorectal cancer: relation to survival. Eur J Surg Oncol 19: 50–60.
Mainiero, F., Pepe, A., Wary, K. K., Spinardi, L., Mohammadi, M., Schlessinger, J. & Giancotti, F. G. (1995). Signal transduction by the α6β4 integrin: Distinct β4 subunit sites mediate recruitment of Shc/Grb2 and association with cytoskeleton of hemidesmosomes. EMBO 14: 4470–4481.
Meijne, A. M., Casey, D. M., Feltkamp, C. A. & Roos, E. (1994). Immuno-EM localization of the beta 1 integrin subunit in wet-cleaved fibronectin-adherent fibroblasts. J Cell Sci 107: 1229–1239.
Miyamoto, S., Akiyama, S. K. & Yamada, K. M. (1995). Synergistic roles for receptor occupancy and aggregation in integrin transmembrane function. Science 267: 883–885.
Mooney, D. J., Langer, R. & Ingber, D. E. (1995). Cytoskeletal filament assembly and the control of cell spreading and function by extracellular matrix. J Cell Sci 108: 2311–2320.
Nicolson, G. L. (1988a). Cancer metastasis: tumor cell and host properties important in colonization of specific secondary sites. Biochim Biophys Acta 948: 175–224.
Nicolson, G. L. (1998b). Organ specificity of tumor metastasis: role of preferential adhesion, invasion and growth of malignant cells at specific secondary sites. Cancer Metastasis Rev 7: 143–188.
Nicolson, G. L. (1989). Metastatic tumor cell interactions with endothelium, basement membrane and tissue. Curr Op Cell Biol 1: 1009–1019.
Nicolson, G. L. (1991). Tumor and host molecules important in the organ preference of metastasis. Cancer Biol 2: 143–154.
Nicolson, G. L. (1995). Tumor interactions with the vascular endothelium and their role in cancer metastasis. In Epithelial–mesenchymal interactions in cancer, Goldberg ID, Rosen EM (ed.) Birkhäuser Verlag: Basel 123–156.
Ogawara, M., Inagaki, N., Tsujimura, K., Takai, Y., Sekimata, M., Ha, M. H., Imajoh Ohmi, S., Hirai, S., Ohno, S. & Sugiura, H. (1995). Differential targeting of protein kinase C and CaM kinase II signaling to vimentin. J Cell Biol 131: 1055–1066.
Orian-Rousseau, V., Aberdam, D., Rousselle, P., Messent, A., Gavrilovic, J., Meneguzzi, G., Kedinger, M. & Simon-Assmann, P. (1998). Human colonic cancer cells synthesize and adhere to laminin-5. Their adhesion to laminin-5 involves multiple receptors among which is integrin α2β1. J Cell Sci 111: 1993–2004.
Price, J. E., Daniels, L. M., Campbell, D. E. & Giavazzi, R. (1989). Organ distribution of experimental metastasis of a human colorectal carcinoma injected in nude mice. Clin Exp Metatasis 7: 55–68.
Rabinovitz, I. & Mercurio, A. M. (1996). The integrin α6β4 and the biology of carcinoma. Biochem Cell Biol 74: 811–821.
Sawada, H., Wakabayashi, H., Nawa, A., Mora, E., Cavanaugh, P. G. & Nicolson, G. L. (1996). Differential motility stimulation but not growth stimulation or adhesion of metastatic human colorectal carcinoma cells by target organ-derived liver sinusoidal endothelial cells. Clin Exp Metastasis 14: 308–314.
Schreiner, C., Bauer, J., Margolis, M. & Juliano, R. L. (1991). Expression and role of integrins in adhesion of human colonic carcinoma cells to extracellular matrix components. Clin Exp Metastasis 9: 163–178.
Schwarz, M. A., Schaller, M. D. & Ginsberg, M. H. (1995). Integrins: Emerging paradigms of signal transduction. Annu Rev Cell Dev Biol 11: 549–599.
Seftor, R. E. B., Seftor, E. A., Gehlsen, K. R., Steler-Stevenson, W. G., Brown, P. D., Ruoslathi, E. & Hendrix, M. J. C. (1992). Role of the αvβ3 integrin in human melanoma cell invasion. Proc Natl Acad Sci USA 89: 1557–1561.
Seufferlein, T. & Rozengurt, E. (1994). Lysophosphatidic acid stimulates tyrosine phosphorylation of focal adhesion kinase, paxillin and p130. J Biol Chem 269: 9345–9351.
Sharpe, A. H., Chen, L. B., Murphy, J. R. & Fields, B. N. (1980). Specific disruption of vimentin filament organization in monkey kidney CV-1 cells by diphtheria toxin, exotoxin A, and cycloheximide. Proc Natl Acad Sci USA 77: 7267–7271.
Stallmach, A., Von Lampe, B., Matthes, H., Bornhöft, G. & Riecken, E. O. (1992). Diminished expression of integrin adhesion molecules on human colonic epithelial cells during the benign to malignant transformation. Gut 33: 342–346.
Streit, M., Schmidt, R., Hilgenfeld, R. U., Thiel, E. & Kreuser, E. D. (1996). Adhesion receptors in malignant transformation and dissemination of gastrointestinal tumors. Recent Results Cancer Res 142: 19–50.
Takazawa, H. (1995). Association between expression of integrin (VLA-3, VLA-5) and malignancy in human colon-cancer. Nippon Rinsho 53: 1672–1677.
Tang, D. G., Timar, J., Grossi, I. M., Renaud, C., Kimler, V. A., Diglio, C. A., Taylor, J. D. & Honn, K. V. (1993). The lipoxygenase metabolite 12(S)-HETE induces a protein kinase C-dependent cytoskeletal rearrangement and retraction of microvascular endothelial cells. Exp Cell Res 207: 361–375.
Tang, D. G., Diglio, C. A. & Honn, K. V. (1994). Activation of microvascular endothelium by eicosanoid 12(S)-hydroxyeicosatetraenoic acid leads to enhanced tumor cell adhesion via up-regulation of surface expression of αvβ3 integrin: A posttranscriptional, protein kinase C- and cytoskeleton-dependent process. Cancer Res 54: 1119–1129.
Tomson, A. M., Scholma, J., Meijer, B., Koning, J. G., De Jong, K. M. & Van Der Werf, M. (1996). Adhesion properties, intermediate filaments and malignant behavior of head and neck squamous cell carcinoma cells in vitro. Clin Exp Metastasis 14: 501–511.
Varedi, M., Ghahary, A., Scott, P. G. & Tredget, E. E. (1997). Cytoskeleton regulates expression of genes for transforming growth factor-beta 1 and extracellular matrix proteins in dermal fibroblasts. J Cell Physiol 172: 192–199.
Wyatt, T. A., Lincoln, T. M. & Pryzwansky, K. B. (1991). Vimentin is transiently co-localized with and phosphorylated by cyclic GMP-dependent protein kinase in formyl-peptide-stimulated neutrophils. J Biol Chem 266: 21274–21280.
Yamada, K. M. & Geiger, B. (1997). Molecular interactions in cell adhesion complexes. Curr Op Cell Biol 9: 76–85.
Yamada, K. M. & Miyamoto, S. (1995). Integrin transmembrane signaling and cytoskeletal control. Curr Op Cell Biol 7: 681–689.
Yatohgo, T., Izumi, M., Kashiwagi, H. & Hayashi, M. (1988). Novel purification of vitronectin from human plasma by heparin affinity chromatography. Cell Struc Funct 13: 281–292.
Young, M. R., Charboneau, S., Lozano, Y., Djordjevic, A. & Young, M. E. (1994). Activation of the protein kinase a signal transduction pathway by granulocyte-macrophage colony-stimulating factor or by genetic manipulation reduces cytoskeletal organization in Lewis lung carcinoma variants. Int J Cancer 56: 446–451.
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Haier, J., Nasralla, M. & Nicolson, G. Different adhesion properties of highly and poorly metastatic HT-29 colon carcinoma cells with extracellular matrix components: role of integrin expression and cytoskeletal components. Br J Cancer 80, 1867–1874 (1999). https://doi.org/10.1038/sj.bjc.6690614
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DOI: https://doi.org/10.1038/sj.bjc.6690614
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