Transcriptional Regulation of Cyclooxygenase-2 in Mouse Skin Carcinoma Cells

Abstract

Many studies have suggested that overexpression of cyclooxygenase-2 (COX-2) contributes to the develop- ment of tumors in several tissues. COX-2 expression tends to be up-regulated in various types of tumors and transformed cell lines, and the overexpression of COX-2 is caused by enhanced transcription of the gene. In an attempt to characterize the signaling pathway leading to the overexpression of COX-2 in the mouse skin carci- noma cell line JWF2, we investigated cis- and trans- acting factors required for COX-2 expression and dem- onstrated a molecular mechanism by which COX-2 is expressed differentially in normal and neoplastic tis- sues. Two regions of the COX-2 promoter containing an E-box and nuclear factor IL6 site were identified as the positive regulatory elements through transient trans- fections with luciferase reporter vectors containing the various 5'-flanking regions of the promoter. Moreover, electrophoretic mobility shift assays and cotransfection experiments showed that upstream stimulatory factors and CCAAT/enhancer-binding proteins (C/EBPs) bind to the E-box and nuclear factor IL6 site, respectively, and functionally transactivate the COX-2 promoter. We also found that C/EBP isoforms are expressed differentially during mouse skin carcinogenesis, suggesting that over- expression of COX-2 in tumors may be caused by a change in C/EBP expression levels.