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J. Biol. Chem., Vol. 277, Issue 47, 44638-44650, November 22, 2002
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From the Important autocrine/paracrine
functions for the adenine nucleotides have been proposed in several
tissues. We addressed the possibility that extracellular ATP would
modulate/mediate hypoxia-induced adventitial fibroblast growth. Acute
hypoxia (3% O2, 10-60 min) increased extracellular
ATP concentrations in adventitial fibroblasts and in lung microvascular
endothelial cells, and chronic hypoxia (3% O2, 14-30
days) markedly attenuated the rate of extracellular ATP
hydrolysis by ecto-nucleotidase(s). Exogenous ATP stimulated [3H]thymidine incorporation in fibroblasts as did UTP,
ADP
Extracellular ATP Is an Autocrine/Paracrine Regulator of
Hypoxia-induced Adventitial Fibroblast Growth
SIGNALING THROUGH EXTRACELLULAR SIGNAL-REGULATED KINASE-1/2 AND
THE Egr-1 TRANSCRIPTION FACTOR*
§,,, and
Developmental Lung Biology Research
Laboratory, Department of Pediatrics, University of Colorado Health
Sciences Center, Denver, Colorado 80262 and ¶ Department of
Pediatrics, National Jewish Medical and Research Center,
Denver, Colorado 80206
, 2-methylthioadenosine triphosphate, adenosine
5'-(
,-methylene)triphosphate, and benzoylbenzoyl-ATP (2'-3'-O-(4-benzoylbenzoyl)-ATP), indicating that both P2Y and P2X
purinoceptors can mediate mitogenic responses. Suramin (100 µM), Cibacron blue 3GA (100 µM), and
pyridoxalphosphate-6-azophenyl-2',-4'-disulfonic acid (100 µM) as well as apyrase (5 units/ml) attenuated hypoxia- and ATP-induced and DNA synthesis, indicating activation and a functional role of purinoceptors under hypoxic conditions. ATP-induced DNA synthesis was augmented by hypoxia in an additive fashion, whereas
ATP and hypoxia synergistically increased growth factor-induced DNA
synthesis, again suggesting that ATP and hypoxia utilize similar signaling pathways to induce proliferation. Indeed, we found that ATP
(100 µM) and hypoxia (3% O2) induced
expression and activation of Egr-1 transcription factor, and both
stimuli acted, in part, through a
Gi/ERK1/2-dependent signaling
pathway. Suramin, Cibacron blue 3GA, and apyrase attenuated
hypoxia-induced ERK1/2 activation and Egr-1 expression. We conclude
that hypoxia induces ATP release from endothelial cells and fibroblasts
and that the activation of P2 purinoceptors is involved in the
regulation of DNA synthesis by fibroblasts under hypoxic conditions.
*
This work was supported by Specialized Center for Research
in Atherosclerosis Grant HL 56481 and National Institutes of Health Grant HL 14985. Preliminary results were presented at the American Thoracic Society Annual Meeting, May 18-23, 2001, San Francisco and
the 41st American Society for Cell Biology Annual Meeting, Dec.
8-12, 2001, Washington, D. C.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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