Regulation of gene expression of a binding protein for fibroblast growth factors by retinoic acid.

Retinoids are potent regulators of growth and differentiation and have shown promise as chemotherapeutic agents against selected cancers in particular squamous cell carcinoma (SCC). Earlier studies from our laboratory showed that a secreted binding protein for fibroblast growth factors (BP) is expressed at high levels in SCC cell lines and tissue samples. Here we investigate whether retinoids affect BP gene expression in SCC. In six different human SCC cell lines, we found that all-trans-retinoic acid (tRA) down-regulated BP mRNA by 39-89% within 24 h. From this group of cell lines, we selected the ME-180 cell line for more detailed studies of the mechanisms of this regulation. tRA down-regulated BP mRNA in a time- and dose-dependent manner. The effect of tRA was reversible, and BP mRNA returned to control levels within 24 h after removal of tRA. We also measured BP mRNA half-life and performed nuclear run-on experiments to study if tRA down-regulates BP by destabilizing the mRNA and/or by decreasing the rate of transcription. BP mRNA in ME-180 cells is very stable with a half-life of >16 h, and tRA decreased BP mRNA with a half-time of 5 h. Actinomycin D and cycloheximide blocked the tRA effect, suggesting that transcriptional regulation as well as de novo protein synthesis contribute to this post-transcriptional regulation of BP mRNA levels. In addition, tRA decreased the rate of BP gene transcription by 2- to 3-fold within 1 h. We conclude that retinoids down-regulate BP gene expression by post-transcriptional as well as by transcriptional mechanisms.