TY - JOUR T1 - Analysis of the Dissociated Steroid RU24858 Does Not Exclude a Role for Inducible Genes in the Anti-Inflammatory Actions of Glucocorticoids JF - Molecular Pharmacology JO - Mol Pharmacol SP - 2084 LP - 2095 DO - 10.1124/mol.106.025841 VL - 70 IS - 6 AU - Joanna E. Chivers AU - Wei Gong AU - Elizabeth M. King AU - Joachim Seybold AU - Judith C. Mak AU - Louise E. Donnelly AU - Neil S. Holden AU - Robert Newton Y1 - 2006/12/01 UR - http://molpharm.aspetjournals.org/content/70/6/2084.abstract N2 - Although repression of inflammatory gene expression makes glucocorticoids powerful anti-inflammatory agents, side effects limit usage and drive the search for improved glucocorticoid receptor (GR) ligands. In A549 pulmonary cells, dexamethasone and the prototypical dissociated ligand RU24858 (Mol Endocrinol11:1245-1255, 1997) repress interleukin (IL)-1β-induced expression of cyclooxygenase (COX)-2 and IL-8. Although RU24858 is a weaker GR ligand, both glucocorticoids showed similar efficacies on transrepression of nuclear factor κB (NF-κB)-dependent transcription, whereas RU24858 yielded less than 12% of the response to dexamethasone on a classic glucocorticoid response element (GRE) reporter (transactivation). Modest NF-κB-dependent transrepression (∼40%), along with analysis of IL-8 transcription rate and the accumulation of unspliced nuclear RNA, indicates that transrepression does not fully account for the repression of genes such as IL-8. This was confirmed by the finding that mRNA degradation is increased by both dexamethasone and RU24858. Analysis of IL-1β-induced steady-state mRNA levels for IL-8 and COX-2 show that dexamethasone- and RU24858-dependent repression of these genes is attenuated by inhibitors of transcription and protein synthesis. Because similar effects were observed with respect to COX-2 and IL-8 protein expression, we conclude that glucocorticoid-dependent gene expression is necessary for repression by both glucocorticoids. Despite RU24858 being defective at classic GRE-dependent transactivation, both dexamethasone and RU24858 induced the expression of potentially anti-inflammatory genes and metabolic genes, suggesting the importance of nontraditional glucocorticoid-dependent gene expression. Thus, classic transactivation- and transrepressionbased screens for anti-inflammatory “dissociated” GR ligands may be flawed because they may not reflect the effects on real glucocorticoid-inducible genes. The American Society for Pharmacology and Experimental Therapeutics ER -