Agonist-specific modulation of glucocorticoid receptor-mediated transcription by immunosuppressants.
Although the immunosuppressive drugs FK506, rapamycin and cyclosporin A have been reported to potentiate transcriptional activation mediated by a non-saturating concentration of the glucocorticoid receptor agonist dexamethasone, the precise mechanism(s) underlying these responses remains unclear. The murine L-929-derived LMCAT cell line stably transfected with the mouse mammary tumor virus promoter-chloramphenicol acetyl transferase reporter gene construct was utilized in the present study to further investigate the mechanism(s) underlying this dexamethasone potentiation as well as the possible agonist specificity of this potentiation. The present data demonstrate that pretreatment (2 h) of LMCAT cells with 10 microM FK506, rapamycin or cyclosporin A results in the potentiation of reporter gene transcription mediated not only by dexamethasone (approximately 12-fold), but also by hydrocortisone (approximately 6-fold) and triamcinolone acetonide (approximately 2.5-fold). In sharp contrast, the data show for the first time that pretreatment with any one of these immunosuppressive drugs suppresses (approximately 2-8-fold) the transcriptional responses mediated by corticosterone, deoxycorticosterone, and cortexolone. Pretreatment of intact LMCAT cells with FK506 increases the subsequent whole cell specific binding of [3H]dexamethasone, but does not increase specific cytoplasmic binding when the tritiated agonist is added directly to cytosolic extracts prepared from the pretreated cells. These data suggest that the FK506-mediated potentiation of the transcriptional responses induced by some agonists, like dexamethasone, may be related to the ability of this immunosuppressant to inhibit the membrane-associated multidrug resistance (MDR) P-glycoprotein, which actively extrudes some steroids from cells. Identical pretreatment with FK506 has no detectable effect on the subsequent whole cell specific binding of [3H]corticosterone, a steroid which is not effectively extruded by the MDR pump. Two additional MDR pump inhibitors, verapamil and quinidine, potentiate (30-fold) the dexamethasone-mediated transcriptional response as expected, but have no detectable effects on a corticosterone-mediated transcriptional response. Unlike immunosuppressive drugs, these ion channel blockers do not bind to receptor-associated immunophilins (FK506-binding proteins or cyclophilins). Collectively, these results suggest that immunosuppressants potentiate a dexamethasone-mediated transcriptional response in LMCAT cells by inhibiting efflux of this steroid. In contrast, these drugs appear to suppress a corticosterone-mediated transcriptional response by a different mechanism, perhaps one involving their binding to glucocorticoid receptor-associated immunophilins.
Department of Physiology and Biophysics, College of Medicine, The University of Iowa, Iowa City 52242, USA.
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D000818:Animals; D003345:Corticosterone / Q000494:pharmacology; D003350:Cortodoxone / Q000494:pharmacology; D016572:Cyclosporine / Q000494:pharmacology; D003900:Desoxycorticosterone / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D017930:Genes, Reporter; D005938:Glucocorticoids / Q000494:pharmacology*; D006854:Hydrocortisone / Q000494:pharmacology; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007739:L Cells; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D011090:Polyenes / Q000494:pharmacology; D011802:Quinidine / Q000494:pharmacology; D011965:Receptors, Glucocorticoid / Q000819:agonists / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D020123:Sirolimus; D016559:Tacrolimus / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014222:Triamcinolone Acetonide / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D005938:Glucocorticoids; D007166:Immunosuppressive Agents; D011090:Polyenes; D011965:Receptors, Glucocorticoid; D011993:Recombinant Fusion Proteins; D003900:Desoxycorticosterone; D003907:Dexamethasone; D016572:Cyclosporine; D014700:Verapamil; D008156:Luciferases; D014222:Triamcinolone Acetonide; D011802:Quinidine; D020123:Sirolimus; D003345:Corticosterone; D003350:Cortodoxone; D006854:Hydrocortisone; D016559:Tacrolimus