chondroitin and Chondrosarcoma

Topics: Aging; Animals; Cartilage; Cells, Cultured; Chondroitin; Chondrosarcoma; Fibronectins; Humans; Hyaluronic Acid; Neoplasms, Experimental; Proteoglycans

The hydraulic conductivity of solutions of Swarm rat chondrosarcoma proteoglycan subunit and of chondroitin 4- and 6-sulfate up to concentrations of 80 mg ml-1 have been measured under physiological conditions using sedimentation velocity and membrane ultrafiltration techniques. This study establishes the very high flow resistance of the proteoglycan and that this resistance is due to its constituent chondroitin sulfate chains. We have also demonstrated little difference in the hydraulic conductivity of chondroitin 4-sulfate as compared to chondroitin 6-sulfate. Studies of hydraulic conductivity of chondroitin sulfate and proteoglycan subunit over a range of salt concentrations demonstrate that the chondroitin sulfates exhibit only a small degree of electrolyte dissipation indicating that their constituent charge groups do not significantly contribute to flow resistance at high mechanical pressures. It appears that the shape and conformation of the polysaccharide backbone and its glycosidic linkages are the factors that primarily govern flow resistance. This is also consistent with the fact that hydraulic conductivity of the proteoglycans and chondroitin sulfates is considerably lower than that of its more charged counterpart heparin but has similar values to hyaluronate. Qualitative agreement between sedimentation analysis and ultrafiltration measurements is also established although the latter technique suffers from not knowing over what distance, adjacent to the membrane, ultrafiltration takes place. It is predicted that the proteoglycans will significantly contribute to flow resistance of cartilagenous tissues which confirms the Maroudas correlation that high proteoglycan concentration in cartilage yields high flow resistance. Further, we establish through a comparison of hydraulic conductivity measurements on hyaluronate, desulfated chondroitin sulfate, chondroitin sulfate, and proteoglycan subunit and osmotic pressure measurements of hyaluronate and proteoglycan that the sulfate groups of the chondroitin sulfate chain play only a small role in the net movement of water relative to the proteoglycan.

Topics: Animals; Chemical Phenomena; Chemistry; Chondroitin; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Chondrosarcoma; Mathematics; Models, Theoretical; Proteoglycans; Rats; Solutions; Ultrafiltration

Chondrocytes isolated from the Swarm rat chondrosarcoma were incubated in culture with [1-3H]glucose for 30 min to 8 h. Labeled proteoglycans were isolated, treated with borohydride under alkaline conditions, and the three complex sugar structures purified: N- and O-linked oligosaccharides and chondroitin sulfate chains. The amount of incorporated radioactivity into each component sugar was analyzed by HPLC after enzyme digestion and hydrolysis. The kinetic data for labeling of each sugar over the time course of the experiment were fit to first-order rate equations and the half times (t1/2) to linear labeling were calculated. The t1/2 values were essentially the same, 5-8 min, for galactose in all three complex sugar structures and for chain glucuronic acid in chondroitin sulfate, while that for xylitol in chondroitin sulfate, 15.8 min, was significantly longer. Thus, oligosaccharide synthesis is concomitant with chondroitin sulfate chain synthesis; the addition of the chondroitin sulfate linkage galactose occurs at or nearly at the same time as chain elongation while the addition of linkage xylose residues to the core protein may precede chain synthesis by up to 8 min. Since the intracellular t1/2 of the core protein precursor for these cells is 45 to 90 min, the data strongly suggest that the addition of xylose is not completed to any significant extent while the polypeptide is still nascent or shortly after its release into the rough endoplasmic reticulum. It is proposed that the addition of xylose to the core protein precursor is a late endoplasmic reticulum or early Golgi event. The analytical data were consistent with the presence of ester phosphate on about 80% of the xylose residues of the newly synthesized proteoglycan.

Topics: Aggrecans; Animals; Cells, Cultured; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Chromatography; Extracellular Matrix Proteins; Glycoproteins; Kinetics; Lectins, C-Type; Oligosaccharides; Protein Processing, Post-Translational; Proteoglycans; Rats

Proteoglycans isolated from the Swarm rat chondrosarcoma were shown to contain 35 mol of phosphate/mol of proteoglycan. While 20% of this phosphate was released by digestion with dilute alkali in the presence of sodium borohydride and is presumably of the phosphoserine/phosphothreonine type, 78% of the phosphate copurified with the peptide-free chondroitin sulfate chains. When chondroitin sulfate chains purified by ethanol precipitation or Sephacryl S200 column chromatography were digested with chondroitinase AC and the digests chromatographed on Bio-Gel P-4, the phosphate co-migrated with a carbohydrate fragment that contained 2 glucuronic acid (one as delta 4,5-unsaturated sugar), 1-galactosamine, 2-galactose, and 1-phosphate residue/xylitol. A second fragment of similar composition but lacking phosphate was also recovered in a ratio of about 3 to 1 relative to the phosphorylated fragment. The phosphate in the chondroitin sulfate linkage region fragment had the alkaline phosphatase sensitivity as well as 31P NMR spectra of a monophosphate esterified to a secondary sugar alcohol. The phosphate was localized on the C-2 of the chain initiating xylose since these residues as xylitol showed a delayed release during acid hydrolysis and the xylitol was recovered intact after periodate oxidation. In the chondrosarcoma, 2-phosphoxylose appears to be a normal synthetic product since [32P]phosphate was readily incorporated into the proteoglycan and the incorporated isotope had similar biochemical properties as the unlabeled phosphate.

Topics: Animals; Carbohydrate Conformation; Carbohydrate Sequence; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Kinetics; Magnetic Resonance Spectroscopy; Oligosaccharides; Phosphorylation; Proteoglycans; Rats

The expression and core protein structure of two proteoglycans, the major cartilage proteoglycan isolated from a rat chondrosarcoma and a small molecular weight chondroitin sulfate proteoglycan isolated from a rat yolk sac tumor, have been compared. The cartilage proteoglycan was not detectable in the cartilage tissue of cartilage matrix deficient (cmd/cmd) neonatal mice by immunofluorescence, but the cmd cartilage did react with antibodies against the core protein of the yolk sac tumor proteoglycan. Radioimmunoassays showed that the core proteins of these proteoglycans are not cross-reactive with each other. Analysis of the core proteins by sodium dodecyl sulfate/polyacrylamide gel electrophoresis after chondroitinase ABC treatment of the proteoglycan revealed a large difference in their sizes. The cartilage proteoglycan core protein had a molecular weight of about 200,000 while the yolk sac tumor proteoglycan core protein migrated with an apparent molecular weight of about 20,000. In addition, the cultured yolk sac tumor cells that make the small proteoglycan did not react with antiserum against the cartilage proteoglycan. These results indicate that the proteoglycan isolated from the yolk sac tumor is similar to the small chondroitin sulfate proteoglycan species found in cartilage and support the existence of at least two dissimilar and genetically independent chondroitin sulfate proteoglycan core proteins.

Topics: Animals; Cartilage; Chondroitin; Chondroitin Lyases; Chondroitin Sulfates; Chondrosarcoma; Cross Reactions; Fluorescent Antibody Technique; Mesonephroma; Mice; Molecular Weight; Radioimmunoassay; Rats; Tissue Distribution

The first case of a myxoid chondrosarcoma, originating in the synovia of the joint and tendon sheaths of the ankle in a 37-year-old man, is reported. Light microscopically the tumor was similar to extraskeletal myxoid chondrosarcoma as previously described. The chondromatous nature of the tumor was further indicated by the histochemical analysis, which revealed the presence of chondroitin 4- and 6-sulphate in the tumor matrix. Ultrastructurally, chondroblast-like cells with dilated rough endoplasmic reticulum, abundant glycogen, some lipid droplets and numerous microvilli-like projections of the cytoplasm dominated. Also mitochondria-rich cells were observed as well as intermediate cell forms. Based on the observations in this case, the frequent occurrence of synovial chondromatosis, the assumed synovial origin of extraskeletal chondromas and the wide morphologic and functional spectrum synovial intima cells may cover, it is suggested that the myxoid chondrosarcoma originated from the synovial intima cells.

Topics: Adult; Ankle Joint; Cell Nucleus; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Cytoplasm; Endoplasmic Reticulum; Histocytochemistry; Humans; Joint Diseases; Male; Microvilli; Synovial Membrane

Chondroitin sulphate synthesis on proteoglycans was decreased in rat chondrosarcoma cell cultures in the presence of cycloheximide (0.1-1.0 muM) or p-nitrophenyl beta-D-xyloside (50 microM). In the presence of cycloheximide the proteoglycan monomer was of larger size, the chondroitin sulphate chains were increased in length, but a similar number of chains was attached to each proteoglycan and the size of the core protein was unaltered. In the presence of p-nitrophenyl beta-D-xyloside (50 microM), chondroitin sulphate synthesis was increased (by 60-80%), but the incorporation into proteoglycans was decreased (by 70%). The chondroitin sulphate chains were of shorter length than in control cultured and the number of chains attached to each proteoglycan was decreased. In cultures with cycloheximide or actinomycin D the synthesis of chondroitin sulphate was less inhibited on beta-xyloside than on endogenous proteoglycan. When the rate of chondroitin sulphate synthesis was decreased by lowering the temperature of cultures, the chains synthesized at 22 and 4 degrees C were much longer than at 37 degrees C, but in the presence of p-nitrophenyl beta-D-xyloside the chains were of the same length at all three temperatures. A model of chain elongation is thus proposed in which the rate of chain synthesis is determined by the concentration of xylosyl acceptor and the length of the chains is determined by the ratio of elongation activity to xylosyl-acceptor concentration.

Topics: Animals; Cartilage; Cells, Cultured; Chemical Phenomena; Chemistry; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Cycloheximide; Dactinomycin; Glycosides; Proteoglycans; Rats; Temperature

Topics: Animals; Carbohydrates; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Fluorides; Kinetics; Neoplasms, Experimental; Pentosyltransferases; Proteoglycans; Rats; Xylose

Topics: Animals; Cells, Cultured; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Enzyme Activation; Glycosaminoglycans; Polysaccharides; Proteoglycans; Rats; Sepharose

Topics: Animals; Bone and Bones; Cartilage; Cations, Divalent; Chickens; Chondroitin; Chondroitin Sulfates; Chondrosarcoma; Galactosyltransferases; Glucuronosyltransferase; Glycosaminoglycans; Hydrogen-Ion Concentration; Kinetics; Neoplasms, Experimental; Pentosyltransferases; Proteoglycans; Rats; Uridine Diphosphate Xylose

Topics: Adult; Blood; Bone Neoplasms; Chondroitin; Chondrosarcoma; Erythropoiesis; Female; Growth Hormone; Humans; Injections, Intravenous; Radiation Effects; Radiotherapy Dosage; Sacrum; Sulfur; Sulfur Radioisotopes

Two mucopolysaccharides, chondroitin 4-sulfate (97.8%) and hyaluronic acid (1.2%), were isolated after exhaustive proteolysis of a transplantable chondrosarcoma of the rat. The chondroitin 4-sulfate was fractionated into three fractions of varying degrees of sulfation and chain length. Keratan sulfate and chondroitin 6-sulfate were absent. Extraction of the fresh tumor gave two protein-polysaccharides of similar carbohydrate composition, one soluble in 0.5 M NaCl, the other insoluble. The latter was solubilized in 4 M guanidine.HCl. A dialyzable fraction from the 4 M guanidine solution may be responsible for the insolubility. Both protein-polysaccharides were antigenic and cross-reacted with similar fractions of bovine and human cartilage.

Topics: Animals; Chondroitin; Chondrosarcoma; Chromatography, Gel; Cross Reactions; Glycosaminoglycans; Guanidines; Hexosamines; Hyaluronic Acid; Hyaluronoglucosaminidase; Male; Mucoproteins; Neoplasm Proteins; Neoplasm Transplantation; Rats; Sarcoma, Experimental; Solubility; Testis; Uronic Acids; Xylose

Topics: Autoradiography; Bone Neoplasms; Chondroitin; Chondrosarcoma; Glycosaminoglycans; Humans; Sulfur Isotopes; Sulfuric Acids

Topics: Bone Neoplasms; Cartilage; Chondroitin; Chondrosarcoma; Chromatography, Paper; Sulfates

Topics: Amino Acids; Animals; Cattle; Chondroitin; Chondrosarcoma; Chromatography; Histocytochemistry; Papain; Peptide Hydrolases; Peptides; Protein Hydrolysates; Research; Serine; Sharks; Spectrophotometry; Sulfates