title: Aldosterone-induced EGFR expression: interaction between the human mineralocorticoid receptor and the human EGFR promoter.
abstract: Aldosterone plays a key role in cardiovascular and renal injury. The underlying mechanisms are not completely understood. Because the epidermal growth factor receptor (EGFR) is involved in the development of fibrosis and vascular dysfunction, upregulation of EGFR expression by aldosterone-bound mineralocorticoid receptor (MR) is an attractive hypothesis. We investigated the effect of aldosterone on EGFR expression in the aorta of adrenalectomized rats and in human aorta smooth muscle cells (HAoSMC) in primary culture. Aldosterone, but not dexamethasone, stimulated EGFR expression in vivo in the aorta as well as in HAoSMC. EGFR degradation was not affected. Aldosterone-induced EGFR expression in HAoSMC was dose dependent and prevented by spironolactone. Furthermore, incubation of HAoSMC with aldosterone led to enhanced EGF-induced ERK1/2 phosphorylation and an EGFR-dependent increase in media fibronectin. EGFR promoter reporter gene assay as well as chromatin immunoprecipitation data indicate that MR interacts with the EGFR promoter. With deletion constructs we gained evidence that this interaction takes place between the hMR and the EGFR promoter regions 316-163 (stronger activation site, EC50 approximately 1.0 nM) and 163-1 (weaker activation site, EC50 approximately 0.7 nM), which do not comprise canonical glucocorticoid response elements and are not activated by the human glucocorticoid receptor. The interactions require in part the NH2-terminal domains of MR. ELISA-based transcription factor DNA binding assay with in vitro synthesized hMR suggest direct binding to region 163-1. Our results indicate that aldosterone leads to enhanced EGFR expression via an interaction with the EGFR promoter, which is MR specific and could contribute to the aldosterone-induced increase in fibronectin abundance.
authors.affiliation: Physiologisches Institut, Universitaet Wuerzburg, Roentgenring 9, 97070 Wuerzburg, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Grossmann
authors.forename: Alexander W
authors.initials: AW
authors.lastname: Krug
authors.forename: Ruth
authors.initials: R
authors.lastname: Freudinger
authors.forename: Sigrid
authors.initials: S
authors.lastname: Mildenberger
authors.forename: Katharina
authors.initials: K
authors.lastname: Voelker
authors.forename: Michael
authors.initials: M
authors.lastname: Gekle
mesh_terms: D000315:Adrenalectomy; D000450:Aldosterone / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D001011:Aorta / Q000378:metabolism*; D002478:Cells, Cultured; D002843:Chromatin; D004247:DNA / Q000378:metabolism; D053938:DNA Fragmentation; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D008024:Ligands; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D018161:Receptors, Mineralocorticoid / Q000235:genetics / Q000378:metabolism*; D017540:Tunica Media / Q000166:cytology / Q000378:metabolism
chemical_list: D002843:Chromatin; D005353:Fibronectins; D008024:Ligands; D018161:Receptors, Mineralocorticoid; D000450:Aldosterone; D004247:DNA; D066246:ErbB Receptors