Abstract
A cell surface-associated adhesive factor (AF) separated from differentiated rat ascites hepatoma AH136B cells (forming cell islands in vivo) has been highly purified by chromatography. AF is assumed to mediate the cell-cell adhesion essential to island formation of the hepatoma cells. A substance, immunologically crossreactive with AF, is present in the ascites fluid or culture medium of the AH136B cells. Because the substance is almost identical to AF in molecular weight and aggregation-promoting activity, it has been concluded that AF is released into the ascites fluid where it is concentrated. Monoclonal antibodies have been raised against AF purified from ascites fluid of AH136B cells. We have obtained a monoclonal antibody, coded MoAF-6D6, that strongly abolishes the aggregation-promoting activity of AF. When AH136B cell islands are incubated in the presence of Fab fragments of MoAF-6D6, cell detachment from the islands is evident within 24 h. Cell islands following 36-h culture show a distinct dissociation and islands completely lose their organization 48 h after culture. The dissociating effect of MoAF-6D6 is neutralized by the addition of AF. These results suggest that AF plays a significant role in the maintenance of cell islands.
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Benchimol S, Fuks A, Jothy S, Beauchemin N, Shirota K, Stanners CP (1989) Carcinoembryonic antigen, a human tumor marker, functions as an intercellular adhesion molecule. Cell 57:327–334
Chen JY, Kurano R, Hirashima M, Hayashi H (1986) An adhesive glycoprotein from rat ascites hepatoma cells potentiates natural cytotoxic activity by rat spleen cells. Immunology 58:95–100
Damsky CH, Knudsen KA, Dorio RJ, Buck CA (1981) Manipulation of cell-cell and cell-substratum interactions in mouse mammary tumor epithelial cells using broad spectrum antisera. J Cell Biol 89:173–184
Damsky CH, Richa DS, Knudsen KA, Buck CA (1983) Identification and purification of a cell surface glycoprotein mediating intercellular adhesion in embryonic and adult tissue. Cell 34:455–466
Duband JL, Dufour S, Hatta K, Takeichi M, Edelman GM, Thiery JP (1987) Adhesion molecules during somitogenesis in the avian embryo. J Cell Biol 104:1361–1374
Fujimoto T, Ogawa K (1983) Cell membrane polarity in dissociated frog urinary bladder epithelial cells. J Histochem Cytochem 31:131–138
Gramzow M, Schröder HC, Uhlenbruck G, Batel R, Müller WEG (1988) Sponge aggregation factor: identification of specific collagen-binding site by means of a monoclonal antibody. J Histochem Cytochem 36:205–212
Hatta K, Okada TS, Takeichi M (1985) A monoclonal antibody disrupting calcium-dependent cell-cell adhesion of brain tissues: possible role of its target antigen in animal pattern formation. Proc Natl Acad Sci USA 82:2789–2793
Hattori R, Ishimaru Y, Kurano R, Hayashi H (1984) An immunomicroscopic study on the distribution of a cell surface-associated adhesive glycoprotein synthesized by a rat ascites hepatoma cells. J Cell Sci 71:95–109
Hayashi H, Ishimaru Y (1981) Morphological and biochemical aspects of adhesiveness and dissociation of cancer cells. Int Rev Cytol 70:139–215
Hifumi M, Hayashi H (1983) Immunological function of adhesive glycoprotein from rat ascites hepatoma cells: production of macrophage chemotactic lymphokine. Immunology 49:245–253
Imhof BA, Vollmers HP, Goodman SL, Birchmeier W (1983) Cell-cell interaction and polarity of epithelial cells: specific perturbation using a monoclonal antibody. Cell 35:667–675
Ishihara H, Ishimaru Y, Hayashi H (1977) Ultrastructural changes of intercellular junctions in rat ascites hepatoma cells with calcium depletion. Br J Cancer 35:643–656
Ishimaru Y, Ishihara H, Hayashi H (1975) An electron microscopic study of tumor cell adhesiveness induced by aggregation promoting factor from rat ascites hepatoma cells. Br J Cancer 31:207–217
Ishimaru Y, Kudo K, Ishihara H, Hayashi H (1976) The induction of tumor cell adhesiveness and intercellular junctions by a glycoprotein of rat ascites hepatoma cell surface. Br J Cancer 34:426–436
Ishimaru Y, Kudo K, Koga Y, Hayashi H (1979) A possible mechanism for island formation by rat ascites hepatoma cells with special reference to the function of aggregation factor at the cell surface. J Cancer Res Clin Oncol 93:123–136
Ishimaru Y, Hattori R, Chen JY, Hayashi H (1985) Cell membrane polarity in rat ascites hepatoma cells: distribution of a cell surface-associated adhesive factor on the cell surfaces. Cell Tissue Res 240:353–359
Kapusta MA, Halberstam D (1964) Preparation of pure 7-S-globulin. Biochim Biophys Acta 93:657–659
Katsuya H, Ishimaru Y, Koono M, Hayashi H (1978) A light and electron microscopic study on complete dissociation of rat ascites hepatoma cells under activation of neutral protease and calcium depletion. Virchows Arch [B] 27:159–172
Koga Y (1981) Electron microscopic study of dissociation of rat ascites hepatoma cells forming islands in vivo. Kumamoto Med J 34:91–107
Kudo K, Tasaki I, Hanaoka Y, Hayashi H (1974) A tumor cell aggregation promoting substance from rat ascites hepatoma cells. Br J Cancer 30:549–559
Kudo K, Hanaoka Y, Hayashi H (1976a) Characterization of tumor cell aggregation promoting substance from rat ascites hepatoma cells: separation of two factors with different properties. Br J Cancer 33:79–90
Kudo K, Hanaoka Y, Hayashi H (1976b) Purification of a tumor cell aggregation-promoting factor associated with rat ascites hepatoma cell surfaces. Br J Cancer 34:88–89
Kurano R (1981) The synthesis of cell surface-associated adhesive factor by rat ascites hepatoma cells and its localization. Kumamoto Med J 34:164–173
Kurano S, Ishida M, Ishimaru Y (1984) Roles of calcium in aggregation of rat ascites hepatoma cells by cell surface-associated adhesive factor. J Cell Sci 66:367–382
Kuratsu J, Yoshinaga M, Hayashi H (1978) Rat lymphocyte mitogenesis by aggregation factor from rat ascites hepatoma cell surface. Br J Cancer 38:224–232
Lowry OH, Rosebrough NJ, Farr AL, Randall RL (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Ocklind C, Odin P, Obrink B (1984) Two different cell adhesion molecules — cell-CAM 105 and a calcium-dependent protein — occur on the surface of rat hepatocytes. Exp Cell Res 151:29–45
Odashima S (1962) Comparative studies on the transplantability of liver cancers induced in rat fed with 3-methyl-4-dimethylamino azobenzen for 3–6 months. Jpn J Cancer Res 53:325–348
Oikawa S, Inuzuka C, Kuroki M, Matsuoka Y, Kosaki G, Nakazato H (1989) Cell adhesion activity of non-specific cross-reacting antigen (NCA) and carcinoembryonic antigen (CEA) expressed on the cell surface: homophilic and heterophilic adhesion. Biochem Biophys Res Commun 164:39–45
Pisam M, Ripoche P (1976) Redistribution of surface macromolecules in dissociated epithelial cells. J Cell Biol 71:907–920
Shibata Y, Ishimaru Y, Kuratsu J (1992) Significance of immunohistochemical localization of an adhesive factor in glioma cells. Kumamoto Med J 43:63–78
Spicer SS, Baron DA, Sato A, Schulte BA (1981) Variability of cell surface glycoconjugates — relation to differences in cell function. J Histochem Cytochem 29:994–1002
Tokuda S (1982) A quantitative analysis of cancer cell aggregation using rat ascites hepatoma cells. Kumamoto Med J 35:131–143
Utsumi S (1969) Stepwise cleavage of rabbit immunoglobulin G by papain and isolation of four types of biologically active Fc fragments. Biochem J 112:343–355
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Ohshima, S., Ishimaru, Y., Honda, M. et al. A monoclonal antibody disrupting cell-cell adhesion of rat ascites hepatoma cells. Cell Tissue Res 273, 363–370 (1993). https://doi.org/10.1007/BF00312839
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DOI: https://doi.org/10.1007/BF00312839