Fibroblast growth factor downregulates expression of a basic helix-loop-helix-type transcription factor, scleraxis, in a chondrocyte-like cell line, TC6.
Scleraxis is a basic helix-loop-helix-type transcription factor that is expressed in sclerotome. Fibroblast growth factor (FGF) is one of the cytokines produced by the cells in skeletal tissues and is a potent modulator of skeletogenesis. The aim of this study was to examine the effects of FGF on the expression of scleraxis in chondrocyte-like cells, TC6. In these cells, scleraxis mRNA was constitutively expressed as a 1 .2 kb message at a high level in contrast to its low levels of expression in fibroblast-like cells or osteoblast-like cells. Upon treatment with FGF, scleraxis mRNA level was decreased within 12 h. This effect was at its nadir at 24 h and the scleraxis mRNA level returned to its base line level by 48 h. The FGF effect was maximal at 1 ng/ml. FGF effects on scleraxis were blocked by actinomycin D but not by cycloheximide, suggesting the involvement of transcriptional events that do not require new protein synthesis. The FGF effects on scleraxis were blocked by genistein, suggesting the involvement of tyrosine kinase in the post-receptor signaling. TGFbeta treatment of TC6 cells enhanced scleraxis mRNA expression; however, combination of the saturation doses of FGF and TGFbeta resulted in suppression of scleraxis mRNA level. BMP2 also suppressed scleraxis mRNA expression in TC6 cells and no further suppression was observed in combination with FGF. These results indicate that scleraxis is expressed in chondrocyte-like TC6 cells and it is one of the targets of FGF action in these cells.
Department of Molecular Pharmacology, Medical Research Institute, Tokyo Medical and Dental University, Japan.
T
T
Kawa-uchi
A
A
Nifuji
N
N
Mataga
E N
EN
Olson
J
J
Bonaventure
K
K
Shinomiya
Y
Y
Liu
M
M
Noda
D051792:Basic Helix-Loop-Helix Proteins; D002460:Cell Line; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects*; D005346:Fibroblast Growth Factors / Q000494:pharmacology*; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics; D017468:Receptors, Fibroblast Growth Factor / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*
D051792:Basic Helix-Loop-Helix Proteins; D005346:Fibroblast Growth Factors; D012333:RNA, Messenger; D017468:Receptors, Fibroblast Growth Factor; D014157:Transcription Factors; C093504:SCX protein, human