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Research ArticleCARDIOVASCULAR

Down-Regulation of Vascular Endothelial Growth Factor Expression after A2A Adenosine Receptor Activation in PC12 Pheochromocytoma Cells

Mark E. Olah and Francine L. Roudabush
Journal of Pharmacology and Experimental Therapeutics June 2000, 293 (3) 779-787;
Mark E. Olah
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Francine L. Roudabush
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Abstract

Vascular endothelial growth factor (VEGF) is an endothelial cell mitogen that promotes angiogenesis during embryonic development and the progression of certain pathologies. This study examined the regulation of VEGF expression by adenosine receptor (AR) activation in PC12 rat pheochromocytoma cells. Treatment of cells with the AR agonist CGS21680 reduced the VEGF mRNA level to ∼20% of that in control cells with an EC50 value of 0.47 nM, indicative of mediation by the A2AAR. Down-regulation of VEGF mRNA by CGS21680 was abolished by pretreatment of cells with the AR antagonist ZM241385. Additionally, ZM241385 alone increased VEGF mRNA by 2.8-fold above basal. RNase protection assays indicated that CGS21680 down-regulated VEGF121, VEGF165, and VEGF189transcripts. VEGF protein secretion was similarly decreased by CGS21680. Under hypoxic conditions, VEGF mRNA expression was reduced by 85.7% after pretreatment with CGS21680. The down-regulation response appears to be mediated predominately by coupling of the A2AAR to Gs because cholera toxin treatment also reduced VEGF expression. The decrease in VEGF mRNA steady-state levels after A2AAR activation is apparently due to a decrease in the VEGF gene transcription rate and not to a decrease in mRNA stability. Thus, depending on the cell type, adenosine may have an inhibitory effect on VEGF production, which may have implications in blood vessel development.

Footnotes

  • Send reprint requests to: Mark E. Olah, P.O. Box 670575, Department of Pharmacology and Cell Biophysics, University of Cincinnati, Cincinnati, OH 45267. E-mail: mark.olah{at}uc.edu

  • ↵1 This study was supported in part by National Institutes of Health/National Cancer Institute Grant RO1 CA79531-01 (to M.E.O.).

  • ↵2 Current address: Duke University Medical Center, Durham, NC 27710.

  • Abbreviations:
    VEGF
    vascular endothelial growth factor
    VEGFR
    vascular endothelial growth factor receptor
    bp
    base pair
    AR
    adenosine receptor
    NECA
    5′-N-ethylcarboxamidoadenosine
    CGS21680
    2-[4-[(2-carboxyethyl)phenyl]ethylamino]-5′-N-ethylcarboxamido-adenosine
    ZM241385
    4-(-2-[7-amino-2-{2-furyl}{1,2,4}triazolo{2,3-a}{1,3,5}triazin-5-yl-amino]ethyl)phenol
    FBS
    fetal bovine serum
    DMSO
    dimethyl sulfoxide
    RPA
    RNase protection assay
    • Received December 20, 1999.
    • Accepted February 21, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 293 (3)
Journal of Pharmacology and Experimental Therapeutics
Vol. 293, Issue 3
1 Jun 2000
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Research ArticleCARDIOVASCULAR

Down-Regulation of Vascular Endothelial Growth Factor Expression after A2A Adenosine Receptor Activation in PC12 Pheochromocytoma Cells

Mark E. Olah and Francine L. Roudabush
Journal of Pharmacology and Experimental Therapeutics June 1, 2000, 293 (3) 779-787;

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Research ArticleCARDIOVASCULAR

Down-Regulation of Vascular Endothelial Growth Factor Expression after A2A Adenosine Receptor Activation in PC12 Pheochromocytoma Cells

Mark E. Olah and Francine L. Roudabush
Journal of Pharmacology and Experimental Therapeutics June 1, 2000, 293 (3) 779-787;
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