Basic fibroblast growth factor inhibits type I collagen gene expression in osteoblastic MC3T3-E1 cells.
We examined the effect of basic fibroblast growth factor (bFGF) on alpha 1(I) procollagen mRNA levels, alpha 1(I) collagen gene transcription, and alpha 1(I) collagen promoter activity in osteoblastic MC3T3-E1 cells. Cells were stably transfected with ColCAT 3.6, containing 3521 base pairs of alpha 1(I) collagen promoter DNA, fused to the CAT reporter gene, or an upstream deletion mutant of ColCAT 3.6 designated ColCAT 2.3. After 48 h, bFGF (0.1-10 nM) inhibited the incorporation of [3H]proline into collagenase-digestible protein (CDP). Indomethacin did not alter the inhibitory effect of bFGF on CDP labeling. Aphidicolin, an inhibitor of DNA synthesis, did not block the inhibitory effect of bFGF on CDP. bFGF (1-10 nM) decreased alpha 1(I) procollagen mRNA levels, with maximal inhibition, nearly 99% of control, caused by 10 nM bFGF. After 48 h, bFGF (1 nM) reduced alpha 1(I) procollagen gene transcription by about 92%. ColCAT 3.6 activity was inhibited with 0.1-10 nM bFGF and was maximally repressed by about 83% with 10 nM bFGF. In contrast, bFGF (1 and 10 nM) caused a stimulation of ColCAT 2.3 activity. These data show that bFGF inhibits collagen synthesis by a transcriptional mechanism and the alpha 1(I) collagen promoter contains DNA sequences which mediate bFGF inhibition of type I collagen gene expression in bone.
Hurley
M M
MM
Department of Medicine, University of Connecticut Health Center, Farmington 06030-1850.
Abreu
C
C
Harrison
J R
JR
Lichtler
A C
AC
Raisz
L G
LG
Kream
B E
BE
D016475:3T3 Cells; D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003094:Collagen / Q000235:genetics*; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D015232:Dinoprostone / Q000096:biosynthesis; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D051379:Mice; D010006:Osteoblasts / Q000378:metabolism*; D011347:Procollagen / Q000235:genetics; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism
D011347:Procollagen; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D016590:Aphidicolin; D003094:Collagen; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D015232:Dinoprostone