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MUCOSAL BIOLOGY

P2X and P2Y purinergic receptors on human intestinal epithelial carcinoma cells: effects of extracellular nucleotides on apoptosis and cell proliferation

¹Autonomic Neuroscience Institute, Royal Free and University College Medical School, London, United Kingdom; ²Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; ³Université Paris 7, Institut Jacques Monod, Paris cedex 5, France; and ⁴School of Natural Sciences, University of California, Merced, California

Submitted 7 May 2004 ; accepted in final form 28 December 2004

Extracellular nucleotides interact with purinergic receptors, which regulate ion transport in a variety of epithelia. With the use of two different human epithelial carcinoma cell lines (HCT8 and Caco-2), we have shown by RT-PCR that the cells express mRNA for P2X₁, P2X₃, P2X₄, P2X₅, P2X₆, P2X₇, P2Y₁, P2Y₂, P2Y₄, P2Y₆, P2Y₁₁, and P2Y₁₂ receptors. Protein expression for P2Y₁ and P2Y₂ receptors was also demonstrated immunohistochemically, and P2X receptor subtype protein was present in the following decreasing order: P2X₄ > P2X₇ > P2X₁ > P2X₃ > P2X₆ > P2X₅ >> P2X₂. The functional presence of P2X₇, P2Y₁, P2Y₂, and P2Y₄ receptors was shown based on the effect of extracellular nucleotides on apoptosis or cell proliferation, and measurement of nucleotide-dependent calcium fluxes using a fluorometric imaging plate reader in the presence of different selective agonists and antagonists. ATP, at high concentrations, induced apoptosis through ligation of P2X₇ and P2Y₁ receptors; conversely, ATP, at lower concentrations, and UTP stimulated proliferation, probably acting via P2Y₂ receptors. We therefore propose that stimulation or dysfunction of purinergic receptors may contribute at least partially to modulation of epithelial carcinoma cell proliferation and apoptosis.

adenosine 5'-triphosphate; purinergic receptors

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