Differential regulation of cyclooxygenase-2 (COX-2) mRNA stability by interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in human in vitro differentiated macrophages

Cyclooxygenase-2 (COX-2) is a highly inducible gene in macrophages by pro-inflammatory cytokines. A major mechanism for cytokine-induced COX-2 expression is stabilization of COX-2 mRNA. In this study, we examined the induction of COX-2 expression by interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) in human primary in vitro differentiated macrophages. IL-1 beta (5 ng/mL) or TNF-alpha (1 ng/mL) induced up to an similar to 40-fold increase of COX-2 mRNA in macrophages during a 2 to 2.5-hr incubation. Run-off experiments demonstrated that cytokine stimulation had only a mild effect on the COX-2 transcription rate (similar to 10-40% increase). The translation blocker cycloheximide (CHM) (10 mg/mL) superinduced COX-2 mRNA during 2 hr of incubation and further stabilized the COX 2 mRNA (T-1/2 > 4 hr). The CHM-superinduced COX-2 mRNA was subject to a rapid degradation after removal of CHM (T-1/2 < 1 hr). Both IL-1 beta and TNF-alpha stabilized cytokine induced COX-2 mRNA (T-1/2 greater than or equal to 2 hr). Maximal stabilization of COX 2 mRNA after a short-term stimulation required the continued presence of IL-1 beta in the medium. Long-term treatment of TNF-alpha destabilized the induced COX-2 mRNA. Cells simultaneously treated with both IL-1 beta and TNF-alpha had a reduced induction of COX-2, IL-1 beta, and IL-6 mRNA. In transcription-arrested cells, the translation blocker puromycin affected the TNF-alpha-induced stabilization and destabilization of COX-2 mRNA, but not the IL-1 beta-induced stabilization. The studies suggest that positive and negative regulation of mRNA stability may play a major role in cytokine mediated COX-2 induction in human macrophages. TNF-alpha map play both pro inflammatory and protective roles during inflammation by regulation of pro-inflammatory gene transcripts. BIOCHEM PHARMACOL 59;2:187-194, 2000. (C) 1999 Elsevier Science Inc.