Long-term exposure to dehydroepiandrosterone affects the transcriptional activity of the glucocorticoid receptor

Although the antightcocorticoid effects of dehydroepiandrosterone (DHEA) have been demonstrated in vivo in many systems, controversial results have been reported by in vitro studies. In order to elucidate the long-term antiglucocorticoid effects of DHEA in vitro in a context more physiological than what proposed by previous works, we set up a system consisting of a carcinoma cell line relying on endogenously produced glucocorticoid receptor (GR) and stably expressing a reporter gene ErbB-2 under the control of a GR-dependent MMTV promoter. These cells grown in presence of low levels of serum glucocorticoids (GC) showed a basal translocation and activity of endogenous GR. The cells reacted to high concentrations of dexamethasone increasing GR nuclear import, although down-regulating receptor expression, and enhancing GR-dependent transcriptional activity, as shown by EMSA assay and expression of the reporter gene ErbB-2. The response to GC was also functional since the increase of ErbB-2 boosted cellular growth. On the contrary, 72 It of incubation with DHEA diminished basal GR-dependent reporter expression and abated cellular proliferation. Analysing molecular mechanisms responsible for this failed transcriptional activity, upon prolonged treatment with DHEA we observed a slow nuclear import of GR not followed by its recruitment to DNA. These data add novel information about the long-term effects of DHEA in vitro. (c) 2006 Elsevier Ltd. All rights reserved.