Opposing effects of fibroblast growth factor and pertussis toxin on alkaline phosphatase, osteopontin, osteocalcin, and type I collagen mRNA levels in ROS 17/2.8 cells.
In rat osteosarcoma (ROS 17/2.8) cells, which express osteoblastic features in culture, basic fibroblast growth factor (bFGF) reduces the level of alkaline phosphatase, type I collagen, and osteocalcin mRNA and increases osteopontin mRNA, independent of growth stimulation. The fibroblast growth factor (FGF) effects are dose dependent (EC50 about 6 pM) and are detected 24 h after addition of the growth factor. bFGF also reduces parathyroid hormone-stimulatable adenylate cyclase and alkaline phosphatase activity in these cells. Concomitant treatment with pertussis toxin (20 ng/ml) opposes the FGF effects. Although cyclic AMP elevating agents mimic pertussis toxin action on some parameters, they produce opposite effects on others, indicating that antagonism between pertussis toxin and bFGF is not mediated by cyclic AMP. bFGF caused a small reduction in steady state NAD-dependent ADP-ribosylation and had no detectable effects on the steady-state levels of the Gi alpha (alpha subunit of the inhibitory G protein) 1, 2, and 3, visualized with specific antibodies in these cells. Although the site of interaction of pertussis toxin and FGF remains to be determined, the findings presented here suggest separate control of growth and differentiation by bFGF and show that pertussis toxin treatment can modulate differentiation in these cells, presumably via Gi proteins.
Rodan
S B
SB
Department of Bone Biology and Osteoporosis Research, Merck Sharp & Dohme Research Laboratory, West Point, Pennsylvania 19486.
Wesolowski
G
G
Yoon
K
K
Rodan
G A
GA
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D037361:Adenylate Cyclase Toxin; D000469:Alkaline Phosphatase / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003094:Collagen / Q000235:genetics*; D000242:Cyclic AMP / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D015675:Osteocalcin / Q000235:genetics*; D053495:Osteopontin; D037342:Pertussis Toxin; D010750:Phosphoproteins / Q000235:genetics; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics*; D012795:Sialoglycoproteins / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D037361:Adenylate Cyclase Toxin; D010750:Phosphoproteins; D012333:RNA, Messenger; D012795:Sialoglycoproteins; C506897:Spp1 protein, rat; D010566:Virulence Factors, Bordetella; D015675:Osteocalcin; D053495:Osteopontin; D011064:Poly Adenosine Diphosphate Ribose; D005346:Fibroblast Growth Factors; D003094:Collagen; D000242:Cyclic AMP; D037342:Pertussis Toxin; D000469:Alkaline Phosphatase; D019204:GTP-Binding Proteins; D015056:1-Methyl-3-isobutylxanthine