Interleukin-1 beta regulates the inducible cyclooxygenase in amnion-derived WISH cells.
The purpose of this study was to determine the mechanism of enhanced prostaglandin synthesis in amnion-derived WISH cell cultures when stimulated by interleukin-1 beta (IL-1 beta). Confluent monolayer cultures of WISH cells were incubated with human recombinant IL-1 beta (0.001-10 ng/ml) for 0-24 hours, while control cells received medium alone. PGE2 production was measured by specific radioimmunoassay. IL-1 beta enhanced the production of PGE2 in a dose- and time-dependent manner with enhanced production detectable by 2 h following exposure. Immunoblot analysis using isoform-specific antibodies showed that the inducible cyclooxygenase enzyme, i.e., COX-2, was expressed by 2 h in IL-1 treated cells, while the constitutive COX-1 remained unaltered in its expression. Northern blot analysis demonstrated that COX-2 mRNA expression was not detected in untreated cells, but became evident after a 30-min exposure to IL-1 beta (10 ng/ml). COX-1 mRNA was detected under basal conditions and did not increase significantly following IL-1 beta treatment. The close parallel between the kinetics of COX-2 mRNA and protein expression and PGE2 accumulation in the medium, as well as the constitutive, unregulated nature of the COX-1 isoform, indicates that cytokine-driven PGE2 formation in WISH cells may be mediated by de novo expression of the novel COX-2 enzyme.
Albert
T J
TJ
Department of Obstetrics and Gynecology, Ohio State University, College of Medicine, Columbus 43210.
Su
H C
HC
Zimmerman
P D
PD
Iams
J D
JD
Kniss
D A
DA
D000650:Amnion / Q000166:cytology* / Q000201:enzymology*; D016718:Arachidonic Acid / Q000494:pharmacology; D002478:Cells, Cultured; D015342:DNA Probes; D015232:Dinoprostone / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D007375:Interleukin-1 / Q000008:administration & dosage / Q000494:pharmacology*; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D007700:Kinetics; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology
D015342:DNA Probes; D007375:Interleukin-1; D007527:Isoenzymes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D016718:Arachidonic Acid; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone