Growth factors increase pericellular proteoglycans independently of their mitogenic effects on A10 rat vascular smooth muscle cells.
Proliferation of vascular smooth muscle cells with the accumulation of proteoglycans in the extracellular matrix is one of the significant changes found in atherosclerotic lesions. In order to clarify the relationship between pericellular proteoglycan and cell growth, we established a simple method for quantitatively estimating the amount of pericellular proteoglycans and investigated the effects of various growth factors on the synthesis of pericellular proteoglycans by cultured A10 rat smooth muscle cells. Analysis of trypsin accessible [35SO4]-labeled material in the pericellular area of the A10 cell culture by Q-sepharose anion-exchange chromatography showed two peaks. One peak, eluted at 0.55 M NaCl, disappeared after treatment with 2 mU/ml of heparitinase, indicating that heparan sulfates (HS) were present. The other peak, which eluted at 0.65 M NaCl, disappeared with 20 mU/ml of chondroitinase ABC, indicating the presence of chondroitin sulfates and dermatan sulfates (CS/DS). We estimated the effects of several growth factors on the synthesis of the pericellular proteoglycans by measuring heparitinase- and chondroitinase-ABC-sensitive radioactivities. Although PDGF-AB significantly stimulated cell proliferation and the synthesis of pericellular CS/DS, its dose-dependent effect on the cell growth did not coincide with that on the proteoglycan synthesis. IGF-I (1 nM) increased pericellular CS/DS but not the cell number, while basic FGF (1 nM) and EGF (1 nM) increased the cell number but not pericellular CS/DS. All the growth factors we examined had no effect on the synthesis of pericellular HS. These results indicate that growth factors increase pericellular proteoglycans independently of their mitogenic effects.
Emoto
N
N
Department of Medicine, Nippon Medical School, Tokyo, Japan. nemoto@nms.ac.jp
Onose
H
H
Yamada
H
H
Minami
S
S
Tsushima
T
T
Wakabayashi
I
I
D000818:Animals; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000378:metabolism / Q000494:pharmacology; D002809:Chondroitin Sulfates / Q000378:metabolism / Q000494:pharmacology; D003871:Dermatan Sulfate / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000494:pharmacology / Q000502:physiology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology / Q000502:physiology; D006133:Growth Substances / Q000494:pharmacology / Q000502:physiology*; D006497:Heparan Sulfate / Q000378:metabolism / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology / Q000502:physiology; D008934:Mitogens / Q000378:metabolism* / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology / Q000502:physiology; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011509:Proteoglycans / Q000032:analysis / Q000096:biosynthesis*; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology
D019765:Chondroitin ABC Lyase; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D004815:Epidermal Growth Factor; D016222:Fibroblast Growth Factor 2; D006133:Growth Substances; D006497:Heparan Sulfate; D007334:Insulin-Like Growth Factor I; D008934:Mitogens; D010982:Platelet-Derived Growth Factor; D011133:Polysaccharide-Lyases; D011509:Proteoglycans; D011994:Recombinant Proteins; C095477:platelet-derived growth factor AB; C021349:heparitinsulfate lyase