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Vol. 59, Issue 3, 493-500, March 2001
-Induced Cyclooxygenase-2 Expression via
Sequential Activation of Ceramide-Dependent Mitogen-Activated Protein
Kinases, and I
B Kinase 1/2 in Human Alveolar Epithelial Cells
Department of Pharmacology, College of Medicine, National Taiwan
University, Taipei, Taiwan
The role of p44/42 mitogen-activated protein kinase (MAPK), p38,
and c-Jun NH2-terminal kinase (JNK) in tumor necrosis
factor (TNF)-
-induced cyclooxygenase (COX)-2 expression was studied in NCI-H292 epithelial cells. TNF--mediated COX-2 expression and
COX-2 promoter activity were inhibited by the MAPK kinase inhibitor
PD98059 or the p38 inhibitor SB203580. Treatment of cells for 10 min
with TNF- resulted in activation of p44/42 MAPK, p38, and JNK.
C2-ceramide (a cell-permeable ceramide analog), bacterial neutral
sphingomyelinase (Smase; an enzyme that degrades sphingomyelin to
ceramide), and N-oleoylethanolamine (a ceramidase inhibitor) all induced activation of MAPKs, COX-2 expression, nuclear
factor (NF)-
B DNA-protein binding, and COX-2 promoter activity. The
inactive analog, dihydro-C2-ceramide, had no effect. SMase- or
C2-ceramide-induced COX-2 expression and COX-2 promoter activity were
also inhibited by PD98059 or SB203580. Glutathione, a neutral SMase
inhibitor, attenuated TNF-- or SMase-induced activation of MAPKs,
COX-2 expression, and COX-2 promoter activity. TNF-- or
C2-ceramide-induced COX-2 promoter activity was inhibited by the
dominant negative mutant of extracellular signal-regulated kinase 2, p38, JNK, IB kinase (IKK)1, or IKK2. IKK activity was stimulated by
either TNF- or C2-ceramide, and these effects were inhibited by
PD98059 or SB203580. All these results suggest that, in NCI-H292
epithelial cells, activation of MAPKs by ceramide contributes to the
TNF- signaling that occurs downstream of neutral SMase activation
and results in the stimulation of IKK1/2, and NF-B in the COX-2
promoter, followed by initiation of COX-2 expression.
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