Retinoic acid regulates insulin-like growth factor-binding protein expression in human osteoblast cells

Retinoic acid (RA) regulates the growth and differentiation of numerous cells types and plays a key role in skeletal development. Previous studies have demonstrated that insulin-like growth factors (IGFs) are important local regulators of bone cell proliferation and differentiation and that IGF-binding proteins (IGFBPs) modulate their activities. In an attempt to test the hypothesis that RA mediates its effects on bone cells in part by regulating IGFBP expression, we first examined the effect of RA on IGFBP expression in human osteoblast model systems and then compared these responses to the effects of RA on IGFBP expression in human skin fibroblasts. The most dramatic effect of RA on IGFBPs++ in all cell types tested was to increase IGFBP-6 messenger RNA (mRNA) abundance more than 1000% of the control value. Significant effects on IGFBP-5 mRNA abundance were also found, with maximal reductions to 35% of control within 24 h of treatment. In addition, RNA maximally increased IGFBP-3 and -4 mRNA to 580% and 390% of the control value, respectively, in SaOS-2 cells, but had variable effects on IGFBP-3 and -4 mRNA levels in human bone cells, U2-OS, and human skin fibroblasts. The levels of the 24-, 29- to 32-, and 38- to 42 kDa IGFBPs in the conditioned medium of RA-treated cultures increased, as determined by ligand blot analysis, whereas the amount of IGFBP-5 was reduced, as determined by RIA. Cycloheximide abolished the RA-stimulated increase in IGFBP-6 mRNA and reduced baseline IGFBP-5 mRNA levels, but did not affect RA-modulated mRNA levels of the other IGFBPs. RA modestly increased the stabilities of all four IGFBP mRNAs, which could contribute to the observed increases in IGFBP-3 and IGFBP-4 mRNA levels; however, the 217% increase in the IGFBP-5 mRNA half- life in the presence of RA could not contribute to the reduction in mRNA levels. In addition, the small increase in the IGFBP-6 mRNA half-life could not account for the 1900% increase in the mRNA level. These data suggest that RA stimulated changes in IGFBP-5 and -6 mRNA levels may in part be mediated by alterations in transcription or other early posttranscription regulatory mechanisms. In conclusion, RA significantly regulates IGFBP expression in human osteoblast cells.