Transcriptional regulation of cyclooxygenase-2 in the human microvascular endothelial cell line, HMEC-1: control by the combinatorial actions of AP2, NF-IL-6 and CRE elements. Mol Cell Biochem 2000 Jan;203(1-2):41-51
Date
03/21/2000Pubmed ID
10724331DOI
10.1023/a:1007045600664Scopus ID
2-s2.0-0034001523 (requires institutional sign-in at Scopus site) 47 CitationsAbstract
Interleukin-1beta (IL-1) is a potent inducer of cyclooxygenase-2 (COX-2) and prostaglandin biosynthesis in many types of cells, yet little is known about the molecular mechanisms regulating IL-1 mediated prostanoid biosynthesis in the endothelium of the microvasculature. Therefore, we examined the cis- and trans-acting factors regulating IL-1-induced COX-2 expression in the human microvascular endothelial cell line, HMEC-1. IL-1 enhanced steady state levels of COX-2 protein and mRNA synthesis by approximately 2-fold which preceded a 2-fold increase in PGF(alpha) biosynthesis. Expression of a series of COX-2 promoter-luciferase constructs in IL-1 treated HMEC-1 cells revealed that the 'full length' (-1432/+59 bp) promoter was 10 times more active than the SV-40 promoter/enhancer and that it could be further activated by IL-1. Surprisingly however, all except for the shortest COX-2 promoter construct retained the ability to respond to IL-1 and luciferase activity driven by -191/+59 bp COX-2 promoter was as responsive to IL-1 as the full-length promoter. Moreover, site-directed promoter mutagenesis and electophoretic mobility shift assays (EMSA) indicate that the combinatorial actions of AP2, NF-IL6, and CRE elements are critical for both constitutive and IL-1-inducible COX-2 promoter activity. Understanding the mechanism(s) regulating COX-2 gene expression and prostaglandin biosynthesis in the microvasculature has important implications with regard to inflammation and angiogenesis in vivo.
Author List
Kirtikara K, Raghow R, Laulederkind SJ, Goorha S, Kanekura T, Ballou LRAuthor
Stanley J. Laulederkind Research Scientist II in the Physiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Base SequenceCCAAT-Enhancer-Binding Protein-delta
CCAAT-Enhancer-Binding Proteins
Cell Line
Cyclic AMP
Cyclooxygenase 2
DNA Primers
DNA-Binding Proteins
Dinoprost
Endothelium, Vascular
Gene Expression Regulation, Enzymologic
Humans
Isoenzymes
Membrane Proteins
Mutation
Nuclear Proteins
Promoter Regions, Genetic
Prostaglandin-Endoperoxide Synthases
Transcription Factor AP-2
Transcription Factors
Transcription, Genetic
