Abstract
The glycosaminoglycans (GAGs) of low (LM) and highly metastatic (HM) cell lines of the Lewis lung tumour (3LL) were compared using [3H]glucosamine labelling techniques. The GAGs isolated from nuclei, cytoplasm, pericellular fractions and medium were analysed by cellulose acetate electrophoresis and by digestion with specific enzymes, and the following conclusions were drawn. 1. Increased cellular uptake and incorporation of [3H]glucosamine into glycoconjugates of the cytoplasm was a typical feature of the highly metastatic cell line after a 48-h labelling. However, there was no elevated radioactivity in glycolipids. 2. Radioactivity of the purified GAGs was two and three times higher in nuclear and cytoplasmic fractions of HM cells than in those of LM cells. There was much less difference between the two cell lines in the pericellular fractions. 3. A definite change from chondroitin sulphate to dermatan sulphate dominancy was recorded in each GAG fraction. Higher heparan sulphate labelling was observed in the cytoplasmic and pericellular GAGs of HM cultures. 4. In the post-labelling period about three times more GAG was present in the extracellular compartment of the HM cultures compared with the LM cultures. 5. In the LM cultures the total GAG-associated radioactivity decreased by 73 per cent in the 48-h chase period whereas in the HM cultures it decreased by only 30 per cent. This indicates a higher rate of GAG degradation in the LM cultures.
Similar content being viewed by others
Abbreviations
- GAG:
-
glycosaminoglycan
- 3LL:
-
Lewis lung tumour
- HM:
-
highly metastatic
- LM:
-
low metastatic
- FCS:
-
foetal calf serum
- PBS:
-
phosphatebuffered saline
- HP:
-
heparin
- HS:
-
heparan sulphate
- HA:
-
hyaluronic acid
- CS:
-
chondroitin sulphate
- DS:
-
dermatan sulphate
- PCA:
-
perchloric acid
- TCA:
-
trichloroacetic acid
- CPC:
-
cetylpyridinium chloride
- E:
-
extracellular
- P:
-
pericellular
- C:
-
cellular
- CY:
-
cytoplasmic
- N:
-
nuclear compartment
- ECM:
-
extracellular matrix
References
Angello, J. C., Danielson, K. G., Anderson, L. W., andHosick, H. L., 1982, Glycosaminoglycan synthesis by subpopulations of epithelial cells from a mammary adenocarcinoma.Cancer Research,42, 2207–2210.
Constantopoulos, G., McComb, R. D., andDekaban, A. S., 1976, Neurochemistry of the mucopolysaccharidoses: brain glycosaminoglycans in normal and four types of mucopolysaccharidoses.Journal of Neurochemistry,26, 901–908.
Doljanski, F., 1982, Cell surface shedding.The Glycoconjugates, Vol. 4, edited by M. I. Horowitz (New York: Academic Press), pp. 155–187.
Fransson, L., Carlstedt, I., andCöster, L., andMalmström, A., 1986, The functions of the heparan sulphate proteoglycans.Functions of the Proteoglycans, Ciba Foundation Symposium 124, edited by D. Evered and J. Whelan (Chichester: Wiley), pp. 125–142.
Gallagher, J. T., Lyon, M., andSteward, W. P., 1986, Structure and function of heparan sulphate proteoglycans.Biochemical Journal,263, 313–325.
Höök, M., Kjellén, L., Johansson, S., andRobinson, J., 1984, Cell surface glycosaminoglycans.Annual Reviews of Biochemistry,53, 847–869.
Iozzo, R. V., 1985, Biology of disease. Proteoglycans: structure, function and role in neoplasia.Laboratory Investigation,53, 373–396.
Irimura, T., Hester, J. E., Yamori, T., Belloni, P. N., Fidler, I. J., Ota, D. M., andNicolson, G. L., 1987, Adhesive properties of human colon carcinoma cells established from primary tumor and metastases.Proceedings of 78th Annual Meeting of AACR,28, 73.
Ishii, K., Katase, A., Andoh, T., andSeno, N., 1982, Inhibition of topoisomerase I by heparin.Biochemical and Biophysical Research Communications,104, 541–547.
Kittlick, P. D., 1985, Glycosaminoglycans. Recent biochemical results in the fields of growth and inflammation.Experimental Pathology, Supplement 10, edited by F. Block, H. David, D. De Paola, C. C. Harriss, R. Hembry, B. A. Lapin, K. Lapis, U. Mohr, Sh. Takayama, G. Waldmann and H. Wrba, (Jena: VEB Gustav Fischer Verlag), pp. 39–45.
Kjellén, L., Oldberg, Å., andHöök, M., 1980, Cell surface heparan sulphate.Journal of Biological Chemistry,255, 10407–10413.
Kraemer, R. J., andCoffey, D. S., 1970, The interaction of natural and synthetic polyanions with mammalian nuclei. I. DNA synthesis.Biochimica et Biophysica Acta,224, 553–567.
Kraemer, R. J., andCoffey, D. S., 1970, The interaction of natural and synthetic polyanions with mammalian nuclei. II. Nuclear swelling.Biochimica et Biophysica Acta,224, 713–717.
Lapis, K., Timár, J., Pál, K., Jeney, A., Timár, F., andKopper, L., 1987, Membrane properties of Lewis lung tumor cells with low and high metastatic capacity. Antimetastatic effect of a glycosaminoglycan biosynthesis blocking agent 5-hexyl-2′-deoxyuridine (HUdR).Cancer Biology and Therapeutics. Proceedings of the 1st Annual H. Lee Moffitt International Symposium, edited by J. G. Cory and A. Szentiványi (New York: Plenum Press), pp. 79–94.
Lapis, K., Timár, J., Timár, F., Pál, K., andKopper, L., 1985, Differences in cell surface characteristics of poorly and highly metastatic Lewis lung tumour variants.Biochemistry and Molecular Genetics of Cancer Metastasis. Proceedings of the NIH Symposium, Bethesda, USA, edited by K. Lapis, L. A. Liotta and A. S. Rabson (Boston: Martinus Nijhoff), pp. 225–235.
Maciag, T., Mehlman, T., Friesel, R., andSchreiber, A. B., 1984, Heparin binds endothelial cell growth factor, the principal endothelial cell mitogen in bovine brain.Science,225, 932–935.
Moczar, E., Laurent, M., andCourtois, Y., 1981, Effect of retinal growth factor and sulfated glycosaminoglycans of bovine lens epithelial cells.Biochimica et Biophysica Acta,675, 132–139.
Newton, D. J., Scott, J. E., andWhiteman, P., 1974, The estimation of acid glycosaminoglycan-Alcian Blue complexes eluted from electrophoretic strrips.Analytical Biochemistry,62, 268–273.
Nicolson, G. L., 1984, Cell surface molecules and tumour metastasis. Regulation of metastasic phenotypic diversity.Experimental Cell Research,150, 3–22.
Ohya, T., andKaneko, Y., 1970, Novel hyaluronidase fromStreptomyces.Biochimica et Biophysica Acta,198, 607–609.
Pál, K., Kopper, L., andLapis, K., 1983, Increased metastatic capacity of Lewis lung tumour cells byin vivo selection procedure.Invasion and Metastasis,3, 174–182.
Pál, K., Kopper, L., Timár, J., Rajnai, J., andLapis, K., 1985, Comparative study on Lewis lung tumour lines with low and high metastatic capacity. I. Growth rate, morphology and resistance to host defence.Invasion and Metastasis,5, 159–169.
Rodén, L., Baker, J. L., Cifonelli, J. A., andMathews, M. B., 1974, Isolation and characterisation of connective tissue polysaccharides.Methods in Enzymology,28, 73–140.
Rollins, B. J., Cathcart, M. K., andCulp, L. A., 1982, Fibronectin-proteoglycan binding as the molecular basis for fibroblast adhesion to extracellular matrix.The Glycoconjugates, Vol. 3, edited by M. I. Horowitz (New York: Academic Press), pp. 289–329.
Saito, H., Yamagata, T., andSuzuki, S., 1969, Enzymatic assay of chondroitin sulphates.Journal of Biological Chemistry,243, 1536–1542.
Schirrmacher, V., andBarz, D., 1986, Characterization of cellular and extracellular plasma membrane vesicles from a low metastatic lymphoma (Eb) and its high metastatic variant (ESb): inhibitory capacity in cell-cell interaction systems.Biochimica et Biophysica Acta,860, 236–242.
Scott, F. J., Fraccastoro, A. P., andTaft, E. B., 1956, Studies in histochemistry: I. Determination of nucleic acids in microgram amounts of tissue.Journal of Histochemistry and Cytochemistry,4, 1–10.
Shing, Y., Folkman, J., Sullivan, R., Butterfield, C., Murray, J., andKlagsbrun, M., 1984, Heparin affinity: purification of a tumor-derived capillary endothelial cell growth factor.Science,223, 1296–1299.
Shivley, J. E., andConrad, H. E., 1976, Formation of anhydrosugars in the depolymerisation of heparan.Biochemistry,15, 3932–3942.
Steck, P. A., Cheong, P. H., Nakajima, M., Yung, W. K. A., Moser, R. P., andNicolson, G. L., 1987, Altered expression of glycosaminoglycans in metastatic 13762NF rat mammary adenocarcinoma cells.Biochemistry,26, 1020–1028.
Timár, J., Moczar, E., Timár, F., Pál, K., Kopper, L., Lapis, K., andJeney, A., 1987, Comparative study on Lewis tumour lines with low and high metastatic capacity. II. Cytochemical and biochemical evidence for differences in glycosaminoglycans.Clinical and Experimental Metastasis,5, 79–87.
Turley, E. A., andTretiak, M., 1985, Glycosaminoglycan production by murine memaloma variantsin vivo andin vitro.Cancer Research,45, 5098–5105.
Wasteson, A., Uthne, K., andWestermark, B., 1973, A novel assay for the biosynthesis of sulphated polysaccharide and its application to studies on the effects of Somatomedin on cultured cells.Biochemical Journal,136, 1069–1074.
Yogeeswaran, G., 1983, Cell surface glycolipids and glycoproteins in malignant transformation.Advances in Cancer Research,38, 289–350.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Pogány, G., Moczar, E., Jeney, A. et al. Comparative study on Lewis lung tumour lines with ‘low’ and ‘high’ metastatic capacity III. Glycosaminoglycan synthesis, transport and degradation in cell lines. Clin Exp Metast 7, 659–669 (1989). https://doi.org/10.1007/BF01753676
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01753676