Techniques used to study regulation of cyclooxygenase-2 promoter sites.
Cyclooxygenase-2 (COX-2), the rate-limiting enzyme for prostaglandin (PG) biosynthesis, plays a key role in inflammation, tumorigenesis, development and circulatory homeostasis. COX-2 expression is rapidly and sensitively regulated by various stimuli, and also its regulation is distinct among cell types at transcriptional and posttranscriptional levels. Therefore, it is important to consider these features of COX-2 expression in the reporter assays we describe in this chapter. Emphasis should be made with regard to two points. Firstly, COX-2 reporter assays should be evaluated by intrinsic COX-2 expression, such as RT-PCR, northern blotting, western blotting, or by PGE(2) measurement. Secondly, one must carefully choose several conditions in the reporter assays for experimental purposes.
Inoue
Hiroyasu
H
Department of Food Science and Nutrition, Faculty of Human Life & Environment, Nara Women's University, Kita-Uoya-Nishimachi, Nara, Japan.
Nakata
Rieko
R
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern / Q000379:methods; D002417:Cattle; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000235:genetics*; D042783:Endothelial Cells / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D005821:Genetic Techniques; D006801:Humans; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014162:Transfection
D051546:Cyclooxygenase 2