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J. Biol. Chem., Vol. 269, Issue 44, 27258-27263, 11, 1994
U Danesch, PC Weber and A Sellmayer
Studying Swiss 3T3 fibroblasts, we report that arachidonic acid strongly
stimulates mRNA levels of the growth-associated immediate early genes c-fos
and Egr-1. Structurally related compounds like arachidonic acid methyl
ester, arachidonyl alcohol, or eicosatetraynoic acid are ineffective,
indicating a specific role of free unesterified arachidonic acid or an
arachidonic acid metabolite in c-fos and Egr-1 mRNA accumulation. Blocking
the conversion of arachidonic acid to prostaglandins by inhibiting
cyclooxygenase abolishes arachidonic acid- induced accumulation of c-fos
and Egr-1 mRNA. Inhibition of the lipoxygenase or cytochrome P-450
epoxygenase pathways has no significant effect on arachidonic acid-induced
c-fos and Egr-1 mRNA levels, indicating that prostaglandin synthesis is
necessary for the increase in c-fos and Egr-1 mRNA. Reversed phase high
performance liquid chromatography revealed prostaglandin E2 (PGE2) as the
major arachidonic acid metabolite in Swiss 3T3 fibroblasts. When added to
the cells, PGE2 stimulates c-fos and Egr-1 mRNA levels to the same degree
as arachidonic acid. Also, the inhibition of arachidonic acid- stimulated
c-fos and Egr-1 mRNA accumulation by indomethacin is reversed by PGE2.
Contrary to reports that PGE2 caused an increase in cAMP levels in Swiss
3T3 fibroblasts, we found that arachidonic acid and PGE2 only minimally
increase cAMP levels as compared with untreated cells. In contrast,
inhibition of protein kinase C by calphostin C and chelerythrine or
down-regulation with phorbol 12-myristate 13-acetate drastically reduces
PGE2 and arachidonic acid-induced c-fos and Egr-1 mRNA levels. These data
indicate that arachidonic acid exerts its stimulatory effect on c-fos and
Egr-1 mRNA via synthesis of PGE2 and subsequent activation of protein
kinase C, probably through a PGE2 receptor coupled to phospholipase C.
Arachidonic acid increases c-fos and Egr-1 mRNA in 3T3 fibroblasts by formation of prostaglandin E2 and activation of protein kinase C
Institut fur Prophylaxe und Epidemiologie der Kreislaufkrankheiten, Klinikum Innenstadt, Universitat Munchen, Federal Republic of Germany.
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