title: Dexamethasone-mediated repression of MUC5AC gene expression in human lung epithelial cells.
abstract: Glucocorticoids regulate gene expression via binding of the ligand-activated glucocorticoid receptor (GR) to glucocorticoid-responsive elements (GRE) in target gene promoters. The MUC5AC gene, which encodes the protein backbone of an abundant secreted airway mucin, has several putative GRE cis-elements in its 5' sequence. Mechanism(s) whereby glucocorticoids regulate mucin genes have not previously been described. In this study, the glucocorticoid dexamethasone (Dex) decreased MUC5AC mRNA abundance in A549 and NCI-H292 cell lines and primary differentiated normal bronchial epithelial cells by 50-80%, suggesting a common mechanism of MUC5AC gene repression in human lung epithelial cells. Kinetic analyses showed that MUC5AC mRNA was not significantly decreased until 6 h after Dex exposure, and that nuclear translocation of GR was biphasic, suggesting that Dex-mediated cis-repression of MUC5AC gene expression was a delayed response of GR translocation. Transfection analyses demonstrated that Dex transcriptionally repressed the MUC5AC promoter. Electrophoretic mobility shift assays with wild-type and mutant oligonucleotide probes showed that GR bound to two GRE cis-sites (nucleotides -930 to -912 and -369 to -351) in the MUC5AC promoter. Analyses of mutated MUC5AC promoter constructs demonstrated that NF-kappaB cis-sites were not involved in Dex-mediated repression of MUC5AC. Dex did not alter mRNA stability of MUC5AC transcripts. Taken together, the data indicate that Dex transcriptionally mediates repression of MUC5AC gene expression in human lung epithelial cells at quiescent states after binding of GR to one or more GRE cis-elements in the MUC5AC promoter.
authors.affiliation: Center for Genetic Medicine Research, Children's Research Institute, Washington, DC 20010, USA.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Chen
authors.forename: Tracey J
authors.initials: TJ
authors.lastname: Nickola
authors.forename: Nancy L
authors.initials: NL
authors.lastname: DiFronzo
authors.forename: Anamaris M
authors.initials: AM
authors.lastname: Colberg-Poley
authors.forename: Mary C
authors.initials: MC
authors.lastname: Rose
mesh_terms: D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology*; D024202:Electrophoretic Mobility Shift Assay; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D008168:Lung / Q000166:cytology; D055271:Mucin 5AC; D009077:Mucins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D016328:NF-kappa B / Q000502:physiology; D011401:Promoter Regions, Genetic; D020871:RNA Stability; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism*
chemical_list: D005938:Glucocorticoids; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009077:Mucins; D016328:NF-kappa B; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone