Life and Medical Sciences
Cell & Developmental Biology
Wiley InterScience Backfile Collection 1832-2000
Chemistry and Pharmacology
Transforming growth factor-β (TGF-β) is present in high levels in bone and plays an important role in osteoblast growth and differentiation. In order to dissect the molecular mechanisms of action of TGF-β on osteoblasts, the effects of TGF-β on the steady state mRNA levels of c-fos, c-jun, and jun-B proto-oncogenes on normal human osteoblast-like cells (hOB) and a transformed human osteoblast cell line (MG-63) were measured. Treatment of hOBs with 2 ng/ml of TGF-β1 resulted in a rapid increase in c-fos mRNA levels as early as 15 min post-treatment. A maximum (10-fold) increase was observed at 30 min after TGF-β treatment followed by a decrease to control values. Similar responses were measured whether the cells were rapidly proliferating or quiescent. TGF-β1 induced jun-B mRNA levels more gradually with steady increase initially observed at 30 min and a maximum induction measured at 2 h post-TGF-β treatment. In contrast, TGF-β treatment caused a time dependent decrease in the c-jun mRNA levels, an opposite pattern to that of jun-B mRNA. Treatment of hOBs with TGF-β1 in the presence of actinomycin-D abolished TGF-β1 induction of c-fos mRNA, suggesting that TGF-β action is mediated via transcription. In the presence of cycloheximide, TGF-β causes super-induction of c-fos mRNA at 30 min, indicating that the c-fos expression by TGF-β is independent of new protein synthesis. Further, transfection of 3 kb upstream region of jun-B promoter linked to a CAT reporter gene into ROS 17/2.8 cells was sufficient to be regulated by TGF-β1. Interestingly, TGF-β treatment also increased the mRNA levels of TGF-β1 itself at 4 h post TGF-β treatment, with a maximum increase observed at 14 h of treatment. TGF-β1 treatment for 30 min were sufficient to cause a delayed increase in TGF-β protein secretion within 24 h. These data support that TGF-β has major effects on hOB cell proto-oncogene expression and that the nuclear proto-oncogenes respond as rapid, early genes in a cascade model of hormone action.
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