Modulation of sulfated glycosaminoglycan and small proteoglycan synthesis by the extracellular matrix.
Cell culture in collagen lattice is known to be a more physiological model than monolayer for studying the regulation of extracellular matrix protein deposition. The synthesis of sulfated glycosaminoglycans (GAG) and dermatan sulfate (DS) proteoglycans by 3 cell strains were studied in confluent monolayers grown on plastic surface, in comparison to fully retracted collagen lattices. Cells were labelled with 35S-sulfate, followed by GAG and proteoglycan analysis by cellulose acetate and SDS-polyacrylamide gel electrophoresis, respectively. The 3 cell strains contracted the lattice in a similar way. In monolayer cultures, the major part of GAG was secreted into culture medium whereas in lattice cultures of dermal fibroblasts and osteosarcoma MG-63 cells but not fibrosarcoma HT-1080 cells, a higher proportion of GAGs, including dermatan sulfate, was retained within the lattices. Small DS proteoglycans, decorin and biglycan, were detected in fibroblasts and MG-63 cultures. They were preferentially trapped within the collagen gel. In retracted lattices, decorin had a higher Mr than in monolayer. Biglycan was detected in monolayer and lattice cultures of MG-63 cells but in lattice cultures only in the case of fibroblasts. In this last case, an up regulation of biglycan mRNA steady state level and down regulation of decorin mRNA was observed, in comparison to monolayers, indicating that collagen can modulate the phenotypical expression of small proteoglycan genes.
Wegrowski
Y
Y
Laboratoire de Biochimie et Biologie Moléculaire, UPRESA CNRS 6021, IFR 53-Biomolécules, Faculté de Médecine, Reims, France.
Gillery
P
P
Kotlarz
G
G
Perreau
C
C
Georges
N
N
Maquart
F X
FX
D000328:Adult; D058578:Biglycan; D015152:Blotting, Northern; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D058575:Decorin; D015536:Down-Regulation; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins; D005347:Fibroblasts / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012516:Osteosarcoma / Q000378:metabolism; D011233:Precipitin Tests; D011509:Proteoglycans / Q000096:biosynthesis* / Q000378:metabolism; D012867:Skin / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism; D013997:Time Factors; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
C546884:BGN protein, human; D058578:Biglycan; C546878:DCN protein, human; D058575:Decorin; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D011509:Proteoglycans; D013455:Sulfur; D003094:Collagen