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Molecular and Cellular Biology, December 2003, p. 9233-9244, Vol. 23, No. 24
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.24.9233-9244.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Transcriptional Regulation of Cyclooxygenase 2 by Bradykinin and Interleukin-1ß in Human Airway Smooth Muscle Cells: Involvement of Different Promoter Elements, Transcription Factors, and Histone H4 Acetylation

Division of Respiratory Medicine, City Hospital, University of Nottingham, Nottingham NG5 1PB, United Kingdom,¹ Department of Pharmacology, National Cardiovascular Centre Research Institute, Suita, Osaka 565-8565, Japan²

Received 11 April 2003/ Returned for modification 16 June 2003/ Accepted 17 September 2003

Bradykinin and interleukin-1ß (IL-1ß) induce cyclooxygenase 2 (COX-2) in human airway smooth muscle cells. Here we extended our study to explore the gene transcriptional regulation. By transfection with various COX-2 promoter reporter constructs, we found that the bp -327-to-+59 promoter region was essential for COX-2 gene transcription by bradykinin and IL-1ß and that the cyclic AMP response element (CRE) was critical in bradykinin-induced transcription, whereas nuclear factor IL-6 and CRE and, to a lesser extent, nuclear factor-B (NF-B) were involved in IL-1ß-induced transcription. An electrophoretic mobility shift assay revealed that both bradykinin and IL-1ß elicited CRE-binding protein-1 (CREB-1) binding, and IL-1ß also elicited CCAAT/enhancer-binding protein ß and NF-B binding to their respective elements in the COX-2 promoter. These transcription factors were associated with the COX-2 promoter, which was dynamically linked to different patterns of histone H4 acetylation by bradykinin and IL-1ß, as demonstrated by chromatin immunoprecipitation. We also revealed that endogenous prostaglandin E₂ was critical in bradykinin-induced COX-2 transcription initiation and involved in IL-1ß-induced COX-2 transcription at a latter stage. Our result provide the first evidence that COX-2 transcriptional regulation by different stimuli involves different promoter elements and transcription factors and is associated with chromatin remodeling after selective histone H4 acetylation in a stimulus-specific manner.

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