Increase in Ref-1 mRNA and protein by thyrotropin in rat thyroid FRTL-5 cells.

Thyrotropin (TSH) induces the expression of fos and jun family genes in thyroid cells. The DNA-binding activity of these gene products (AP-1) has been shown to be enhanced by ubiquitous nuclear redox factor-1 (Ref-1). We thus examined whether TSH regulates Ref-1 gene expression in rat thyroid FRTL-5 cells. Northern blot analysis revealed that the abundance of Ref-1 mRNA significantly increased within 3 hours after TSH followed by a sustained increase until 12 hours. The increase was also induced by treatment with forskolin but not in the presence of cycloheximide, indicating that the TSH effect on Ref-1 mRNA is mediated by intracellular cAMP and requires de novo protein synthesis. Consistent with the elevation of the mRNA level, Western blot analysis showed an increase in Ref-1 protein 3 hours after TSH. The level continued to increase until 12 hours. These results suggested that increased Ref-1 by TSH might regulate the binding activity of AP-1 in thyroid cells. Considering that Ref-1 also has a DNA repair function, Ref-1 may play dual roles in gene regulation and DNA repair processes in thyroid cells.