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J. Biol. Chem., Vol. 268, Issue 29, 21875-21882, Oct, 1993
SE Barbour and EA Dennis
Macrophage-like P388D1 cells release [3H]arachidonic acid and produce
prostaglandin E2 (PGE2) upon stimulation with bacterial lipopolysaccharide
(LPS) and platelet-activating factor (PAF). To determine whether group II
phospholipase A2 (PLA2) is involved in this release, we treated P388D1
cells with antisense inhibitors specific for group II PLA2 RNA. Treatment
with oligonucleotide ASGII decreased PLA2 activity in P388D1 cell
homogenates by approximately 60% and reduced the release of [3H]arachidonic
acid and PGE2 from activated cells to nearly resting cell levels. The
inhibition by antisense oligonucleotide ASGII was blocked when its sense
complement, SGII, was included in the incubation mixture. Stably
transfected P388D1 cells expressing an antisense construct for group II
PLA2 also produced reduced quantities of PGE2 in response to LPS and PAF.
These data suggest that prostaglandin production by activated P388D1 cells
involves phospholipid hydrolysis by group II PLA2. Oligonucleotide ASGII
also blocked the appearance of a heparin-releasable group II PLA2 in the
culture supernatants of P388D1 cells. The disappearance of this protein
correlated with reduced PGE2 production by activated cells, indicating that
an extracellular heparin-associated pool of group II PLA2 is involved in
prostaglandin production by P388D1 cells.
Antisense inhibition of group II phospholipase A2 expression blocks the production of prostaglandin E2 by P388D1 cells
Department of Chemistry, University of California at San Diego, La Jolla 92093-0601.
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