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Department of Physiology and Biophysics, School of Medicine, Case
Western Reserve University, Cleveland, Ohio 44106-4970
HL-60 human promyelocytic leukocytes express G protein-coupled
P2U-purinergic nucleotide receptors (P2UR or
P2Y2R) that activate inositol phospholipid hydrolysis and
Ca2+ mobilization in response to ATP or UTP. We examined
the expression of functional P2UR and P2UR mRNA
levels during in vitro differentiation of HL-60 cells by
dibutyryl-cAMP (Bt2cAMP), which induces a
granulocyte/neutrophil phenotype, or by phorbol-12-myristate-13-acetate
(PMA), which induces a monocyte/macrophage phenotype. Both
P2UR function and P2UR mRNA levels were only
modestly attenuated during granulocytic differentiation by
Bt2cAMP. In contrast, P2UR function, as assayed by either Ca2+ mobilization or inositol trisphosphate
generation, was greatly reduced in PMA-differentiated cells. This
inhibition of P2UR function was strongly correlated with
PMA-induced decreases in P2UR mRNA levels, as assayed by
Northern blot analysis or reverse transcription-polymerase chain
reaction-based quantification. Although PMA induced an early, transient
up-regulation of P2UR mRNA, this was rapidly followed by a
sustained decrease in P2UR mRNA to a level 5-10-fold lower than that in undifferentiated HL-60 cells. The half-life of the P2UR transcript in HL-60 cells was ~60 min, and this was
not affected by acute exposure (
4 hr) to Bt2cAMP or PMA.
PMA down-regulated P2UR mRNA in THP-1 monocytes and HL-60
granulocytes but not in A431 human epithelial cells or human
keratinocytes. P2UR mRNA was also down-regulated in THP-1
monocytes differentiated into inflammatory macrophages by
-interferon and endotoxin. These data indicate that myeloid
leukocytes possess tissue-specific mechanisms for the rapid modulation
of P2UR expression and function during differentiation and
inflammatory activation.
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