concanavalin-a and Chondrosarcoma

SW1353 chondrosarcoma cells cultured in the presence of interleukin-1, concanavalin A or PMA secreted procollagenase 3 (matrix metalloproteinase-13). The enzyme was detected in the culture medium by Western blotting using a specific polyclonal antibody raised against recombinant human procollagenase 3. Oncostatin M enhanced the interleukin-1-induced production of procollagenase 3, whereas interleukin-4 decreased procollagenase 3 synthesis. The enzyme was latent except when the cells had been treated with concanavalin A, when a processed form of 48 kDa, which corresponds to the active form, was found in the culture medium and collagenolytic activity was detected by degradation of 14C-labelled type I collagen. The concanavalin A-induced activation of procollagenase 3 coincided with the processing of progelatinase A (matrix metalloproteinase-2) by the cells, as measured by gelatin zymography. In addition, progelatinase B (matrix metalloproteinase-9) was activated when gelatinase A and collagenase 3 were in their active forms. Concanavalin A treatment of SW1353 cells increased the amount of membrane-type-1 matrix metalloproteinase protein in the cell membranes, suggesting that this membrane-bound enzyme participates in an activation cascade involving collagenase 3 and the gelatinases. This cascade was effectively inhibited by tissue inhibitors of metalloproteinases-2 and -3. Tissue inhibitor of metalloproteinases-1, which is a much weaker inhibitor of membrane-type 1 matrix metalloproteinase than tissue inhibitors of metalloproteinases-2 and -3 [Will, Atkinson, Butler, Smith and Murphy (1996) J. Biol. Chem. 271, 17119-17123], was a weaker inhibitor of the activation cascade.

Topics: Blotting, Western; Cell Membrane; Chondrosarcoma; Collagenases; Concanavalin A; Culture Media, Conditioned; Enzyme Activation; Enzyme Induction; Fibrinolysin; Gelatinases; Humans; Interleukin-1; Matrix Metalloproteinase 13; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Metalloendopeptidases; Protease Inhibitors; Tetradecanoylphorbol Acetate; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinase-2; Tissue Inhibitor of Metalloproteinase-3; Tumor Cells, Cultured

Clear cell chondrosarcoma, a subtype and separate entity from the conventional chondrosarcoma, is characterized by its special histologic features, site of predilection, slow growth and better prognosis. Three cases are presented with elucidation of clinicopathologic correlation and detection by ABC immunohistochemical method using several antibodies. The observation of positive reaction to S-100 protein, vimentin, anti-alpha-chymotrypsin and Lysozyme by the tumor cells of clear cell chondrosarcoma, similar to chondrosarcoma and chondroblastoma, proves that this tumor has its origin from the cartilaginous tissue. It was found for the first time that the clear cell chondrosarcoma was positive for wheat germ agglutinin and concanavalin A. The authors believe that clear cell chondrosarcoma may result from the anaplastic change of chondroblastoma cells into another subtype of that tumor. The osteoblastlike multinucleated giant cells, retaining the antigens of phagocytes, are not considered to be neoplastic.

Topics: Adult; Bone Neoplasms; Chondrosarcoma; Concanavalin A; Female; Femoral Neoplasms; Histocytochemistry; Humans; Immunohistochemistry; Male; Tibia; Wheat Germ Agglutinins

Tunicamycin, an inhibitor of dolichol-diphospho-N-acetylglucosamine formation and hence an inhibitor of N-linked oligosaccharide biosynthesis, suppressed total proteoglycan synthesis by Swarm rat chondrosarcoma chondrocytes without affecting the size of the proteoglycan molecule, its secretion from the cell, or its ability to be retained in the extracellular matrix. In addition, tunicamycin did not substantially alter the ability of the chondrocytes to polymerize glycosaminoglycan onto an exogenous beta-D-xyloside acceptor. A secondary effect of tunicamycin suppression of proteoglycan synthesis was that a lesser amount of newly synthesized proteoglycan diffused from the extracellular matrix into the culture medium. The ability of exogenous hyaluronic acid and proteoglycan to increase the percentage of newly synthesized 35S-proteoglycan in the medium in tunicamycin-treated cultures indicates that matrix retention of 35S-proteoglycan is related to the total extracellular uronic acid content rather than to the presence or absence of mannose oligosaccharides bound to the proteoglycan molecule. These noncytotoxic concentrations of tunicamycin (33-333 ng/ml) decreased [3H]mannose incorporation to the same extent that they decreased total [35S]sulfate and [3H]serine incorporation, and caused the chondrocyte to synthesize and secrete a species of beta-hexosaminidase that was mannose-deficient as assessed by its failure to bind to concanavalin A. The additional finding of decreased insulin binding to tunicamycin-treated chondrosarcoma chondrocytes suggested that the inhibition of proteoglycan synthesis was due to diminution of receptors which respond to stimulatory hormones.

Topics: Acetylglucosaminidase; Animals; Chondrosarcoma; Chromatography, Affinity; Concanavalin A; Glucosamine; Insulin; Kinetics; Proteoglycans; Rats; Receptor, Insulin; Sarcoma, Experimental; Tunicamycin