Summary
To contribute to the investigation of the composition of the extracellular matrix in epithelial tumours, mammary gland tissues of dogs (including tumours, hyperplasias and normal tissue as well as metastatic lesions in lymph nodes and lung) were studied histochemically and immunohistochemically for distribution of sulphated glycosaminoglycans (s-GAGs). The formaline-fixed tissue was stained by alcian blue at pH 5.8, using the ‘critical electrolyte concentration’ to study the degree of sulphation of s-GAGs. s-GAGs were characterized by degradation with enzymes and nitrous acid and by immunohistochemistry with two anti-chondroitin sulphate monoclonal antibodies. The light microscopic investigation of s-GAG deposits revealed a limited number of patterns of their distribution. The main s-GAGs found in the mammary gland tumours of dogs and in metastatic lesions were chondroitin sulphate (CS) and heparin/heparan sulphate (HEP/HS). CS accumulated in diffuse structures between epithelial cells as well as around clusters of tumour cells. The latter pattern, possibly representing a mesenchymal reaction to the tumour, was present in 74% of the tumours, and in 67% of these, highly sulphated CS was present. A diffuse accumulation of CS was present almost exclusively in complex and mixed tumours; because of the expression of the 3B3 epitope for CS in immature cartilage the spindle cells of complex tumours are argued to be the precursors of the cartilage in mixed tumours. HEP/HS was stored mainly in mast cells that were found in increased numbers in hyperplasias and tumours. By pretreatment of microscopic slides with chondroitinase AC or ABC immunostaining of fibronectin could be made possible in areas in which CS was abundantly present, suggesting that CS may mask fibronectin epitopes. It is concluded that CS with different degrees of sulphation is the most important s-GAG in the extracellular matrix of mammary tumours of dogs. CS and other s-GAGs accumulate at different sites and may have a different pathogenetic significance.
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References
Abbas, K. A., Lichtman, A. H. & Pober, J. S. (1991). Cellular and Molecular Immunology, 289 WB Saunders: Philadelphia
Adany, R., Heimer, R., Caterson, B., Sorrell, J. M. & Iozzo, R. V. (1990). Altered expression of chondroitin sulphate proteoglycan in the stroma of human colon carcinoma. Hypomethylation of PG-40 gene correlates with increased PG-40 content and mRNA. J Biol Chem 265: 11389–11396.
Albrechtsen, R., Wewer, U. M. & Liotta, L. A. (1986). Basement membranes in human cancer. Pathol Annu 51: 251–276.
Alini, M. & Losa, G. A. (1991). Partial characterization of proteoglycans isolated from neoplastic and nonneoplastic human breast tissue. Cancer Res 51: 1443–1447.
Andres, J. L., Stanley, K., Cheifetz, S. & Massagué, J. (1989). Membrane-anchored and soluble forms of betaglycan, a polymorphic proteoglycan that binds transforming growth factor-β. J Cell Biol 109: 3137–3145.
Caterson, B., Mahmoodian, F., Sorrell, J. M., Hardingham, T. E., Bayliss, M. T., Carney, S. L., Ratcliffe, A. & Muir, H. (1990). Modulation of native chondroitin sulphate structure in tissue development and in disease. J Cell Sci 97: 411–417.
David, G. (1993). Integral membrane heparan sulfate proteoglycans. FASEB J 7: 1023–1030.
Edward, M., Grant, A. W. & Mackie, R. M. (1992). Human melanoma cell-derived factor(s) stimulate fibroblast glycosaminoglycan synthesis. Int J Cancer 52: 499–503.
Erlinger, R., Schumacher, U. & Welsch, U. (1990). Ultrastructural localisation of glycosaminoglycans in the human mammary gland. Acta Histochem Suppl. XL: 65–70.
Forsberg, L. S., Lazarus, S. C., Seno, N., Devinney, R., Caughey, G. H. & Gold, W. M. (1988). Dog mastocytoma proteoglycans: occurrence of heparin and oversulphated chondroitin sulphates, containing trisulphated disaccharides, in three cell lines. Biochim Biophys Acta 967: 416–428.
Guelstein, V. I., Tchypysheva, T. A., Ermilova, V. D. & Ljubimov, A. V. (1993). Myoepithelial and basement membrane antigens in benign and malignant human breast tumours. Int J Cancer 53: 269–277.
Hampe, J. F. & Misdorp, W. (1974). Tumors and dysplasias of mammary gland. Bull Wld Hlth Org 50: 111–133.
Iozzo, R. V. (1985a). Proteoglycans: structure, function and role in neoplasia. Lab Invest 53: 373–392.
Iozzo, R. V. (1985b). Neoplastic modulation of extracellular matrix: colon carcinoma cells release polypeptides that alter proteoglycan metabolism in colon fibroblasts. J Biol Chem 260: 7464–7473.
Iozzo, R. V. (1987). Proteoglycans and the intercellular tumor matrix. In Current Topics in Pathology, Seifert G Springer: Berlin 207–221.
Iozzo, R. V. (1988). Proteoglycans and neoplasia. Cancer Metast Rev 7: 39–50.
Iozzo, R. V. (1997). The family of the small leucine-rich proteoglycans: key regulators of matrix assembly and cellular growth. Crit Rev Biochem Mol Biol 32: 141–174.
Jackson, R. L., Busch, S. J. & Cardin, A. D. (1991). Glycosaminoglycans: molecular properties, protein interaction and role in physiological processes. Physiol Rev 71: 481–539.
Klebe, R. J., Escobedo, L. V., Bentley, K. L. & Thompson, L. K. (1986). Regulation of cell motility, morphology, and growth by sulphated glycosaminoglycans. Cell Motil 6: 273–281.
Losa, G. A. & Alini, M. (1993). Sulphated proteoglycans in the extracellular matrix of human breast tissues with infiltrating carcinoma. Int J Cancer 54: 552–557.
Merrilees, M. J. & Finlay, G. J. (1985). Human tumor cells in culture stimulate glycosaminoglycan synthesis by human skin fibroblasts. Lab Invest 53: 30–36.
Misdorp, W. & Hart, A. A. M. (1976). Prognostic factors in canine mammary cancer. J Natl Cancer Inst 56: 779–786.
Nara, Y., Kato, Y., Torii, Y., Tsuji, Y., Nakagaki, S., Goto, S., Isobe, H., Nakashima, N. & Takeuchi, J. (1997). Immunohistochemical localization of extracellular matrix components in human breast tumours with special reference to PG-M/versican. Histochem J 29: 21–30.
Nara, Y., Takeuchi, J., Yoshida, K., Fukatsu, T., Nagasaka, T., Kawaguchi, T., Meng, N., Kikuchi, H. & Nakashima, N. (1991). Immunohistochemical characterisation of extracellular matrix components of salivary gland tumors. Br J Cancer 64: 307–314.
Niewold, T. A., Flores Landeira, J. M., Van den Heuvel, LPWJ, Ultee, A., Tooten, P. C. J. & Veerkamp, J. H. (1991). Characterization of proteoglycans and glycosaminoglycans in bovine renal AA-type amyloidosis. Virchows Arch B Cell Pathol Mol Pathol 60: 321–328.
Palmer, T. E. & Monlux, A. W. (1979). Acid mucopolysaccharides in mammary tumors of dogs. Vet Pathol 16: 493–509.
Pulley, L. T. (1973). Ultrastructural and histochemical demonstration of myoepithelium in mixed tumors of the canine mammary gland. Am J Vet Res 34: 1513–1522.
Roche, W. R. (1986). The nature and significance of tumor-associated mastcells. J Pathol 148: 175–182.
Roskelley, C. D. & Bissell, M. J. (1995). Dynamic reciprocity revisited: a continuous, bidirectional flow of information between cells and the extracellular matrix regulates mammary epithelial cell function. Biochem Cell Biol 73: 391–397.
Santra, M., Mann, D. M., Mercer, E. W., Skorski, T., Calabretta, B. & Iozzo, R. V. (1997). Ectopic expression of decorin protein core causes generalized growth suppression in neoplastic cells of various histogenetic origin and requires endogenous p21, an inhibitor of cyclin-dependent kinases. J Clin Invest 100: 149–157.
Scott, J. E. & Dorling, J. (1965). Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemistry 5: 221–233.
Silberstein-GB & Daniel, C. W. (1982). Glycosaminoglycans in the basal lamina and extracellular matrix of developing mouse mammary duct. Dev Biol 90: 215–222.
Takeuchi, J., Sobue, M., Sato, E., Shamoto, M., Suzuki, S. & Kimata, K. (1976). Variation in glycosaminoglycan components of breast tumours. Cancer Res 36: 2133–2139.
Vos, J. H., Van den Ingh, TSGAM, Misdorp, W., Molenbeek, R. F., Van Mil, F. N., Rutteman, G. R., Ivanyi, D. & Ramaekers, F. C. S. (1993a). Immunohistochemistry with keratin, vimentin, desmin and α-smooth muscle actin monoclonal antibodies in canine mammary gland: benign mammary tumours and ductectasias. Vet Quart 15: 89–95.
Vos, J. H., Van den Ingh, TSGAM, Misdorp, W., Molenbeek, R. F., Van Mil, F. N., Rutteman, G. R., Ivanyi, D. & Ramaekers, F. C. S. (1993b). Immunohistochemistry with keratin, vimentin, desmin and α-smooth muscle actin monoclonal antibodies in canine mammary gland: malignant mammary tumours. Vet Quart 15: 96–101.
Yamagata, M., Suzuki, S., Akiyama, S. K., Yamada, M. & Kimata, K. (1989). Regulation of cell-substrate adhesion by proteoglycans immobilized on extracellular substrates. J Biol Chem 264: 8012–8018.
Yamori T., Ota, D. M., Cleary, K. R. & Irimura, T. (1988). Increased content of chondroitin sulphate proteoglycan in human colorectal carcinoma metastases compared with the primary tumor as determined by an anti-chondroitin-sulphate monoclonal antibody. J Cell Biochem 36: 405–416.
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Hinrichs, U., Rutteman, G. & Nederbragt, H. Stromal accumulation of chondroitin sulphate in mammary tumours of dogs. Br J Cancer 80, 1359–1365 (1999). https://doi.org/10.1038/sj.bjc.6690529
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DOI: https://doi.org/10.1038/sj.bjc.6690529
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