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and
PMA in human monocytic U-937 cells
1 Combined Program in Pulmonary
and Critical Care Medicine,
Regulation of eotaxin expression was
investigated in U-937 cells, a human monocyte-like cell line. Eotaxin
mRNA was induced by tumor necrosis factor-
(TNF-
; 0.1-100
ng/ml) and phorbol 12-myristate 13-acetate (PMA; 0.01-1 µM).
PMA-induced eotaxin mRNA expression was of greater magnitude and was
maximal at a later time point than TNF-
-induced expression (16 h vs.
2 h after stimulation), which was consistent with eotaxin protein
expression detected by immunocytochemistry. Dexamethasone (0.01-10
µM) decreased eotaxin mRNA expression in both TNF-
- and
PMA-stimulated U-937 cells. PMA-induced eotaxin mRNA expression was
inhibited by cycloheximide (10 µg/ml), whereas TNF-
-induced
expression was not. The protein kinase C (PKC) inhibitor staurosporine
(10-50 nM) inhibited PMA-induced eotaxin mRNA expression, whereas
TNF-
-induced expression was enhanced by this reagent. These results
suggest that eotaxin expression can be induced by more than one
mechanism: the PMA-triggered pathway is mediated by PKC activation and
requires new protein synthesis, whereas the TNF-
-triggered pathway
is independent of PKC and protein synthesis. TNF-
- and PMA-induced
pathways are both associated with nuclear factor-
B, because its
binding activity was enhanced in the presence of these stimuli, and
both pathways were limited by its inhibitor, diethyldithiocarbamate.
dexamethasone; protein kinase C; nuclear factor-
B; cycloheximide
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