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Cell and Tissue Research

, Volume 321, Issue 3, pp 411–418 | Cite as

Human melanomas express functional P2X7 receptors

  • Nicholas White
  • Peter E. M. Butler
  • Geoffrey BurnstockEmail author
Regular Article

Abstract

Adenosine 5′-triphosphate is known to function as a potent extracellular messenger, producing its effects via a distinct family of cell surface receptors. Different receptor subtypes have been shown to modulate different cellular functions such as proliferation, differentiation and apoptosis. We have investigated the functional expression and apoptotic action of the P2X7 receptor in human malignant melanoma tissue and cells. Incubation of cells with the potent P2X7 receptor agonist 2′–3′-O-(4-benzoyl-benzoyl) adenosine 5′-triphosphate leads to a decrease in cell number, which is dose-dependent and reversible by the antagonist 1-N,O-bis-[5-isoquinoline-sulfonyl]-N-methyl-L-tyrosyl)-4-phenyl-piperazine. Synthesis of the P2X7 receptor by these cells has been established by reverse transcriptase-polymerase chain reaction, immunohistochemistry, immunocytochemistry and cellular accumulation of the fluorescent DNA-binding dye YO-PRO-1. The P2X7 receptors have been shown to mediate apoptotic actions of extracellular nucleotides and represent a novel target for melanoma therapy.

Keywords

Purinergic receptors ATP P2X7 Melanoma Cancer Human 

Notes

Acknowledgements

The P2X7 receptor antibody was a gift from Roche Palo Alto (Palo Alto, Calif., USA).

References

  1. Abbracchio MP, Burnstock G (1994) Purinoceptors: are there families of P2X and P2Y purinoceptors? Pharmacol Ther 64:445–475CrossRefPubMedGoogle Scholar
  2. Abraham EH, Salikhova AY, Rapaport E (2003) ATP in the treatment of advanced cancer. In: Schwiebert EM (ed) Extracellular nucleotides and nucleosides. Elsevier Science, San Diego, pp 415–452Google Scholar
  3. Adrian K, Bernhard MK, Breitinger HG, Ogilvie A (2000) Expression of purinergic receptors (ionotropic P2X1–7 and metabotropic P2Y1–11) during myeloid differentiation of HL60 cells. Biochim Biophys Acta 1492:127–138PubMedGoogle Scholar
  4. Agteresch HJ, Dagnelie PC, van der Gaast A, Stijnen T, Wilson JH (2000) Randomized clinical trial of adenosine 5′-triphosphate in patients with advanced non-small-cell lung cancer. J Natl Cancer Inst 92:321–328CrossRefPubMedGoogle Scholar
  5. Burnstock G (1998) Sympathetic purinergic transmission in small blood vessels. Trends Pharmacol Sci 9:116–117CrossRefGoogle Scholar
  6. Burnstock G, Dumsday B, Smythe A (1972) Atropine resistant excitation of the urinary bladder: the possibility of transmission via nerves releasing a purine nucleotide. Br J Pharmacol 44:451–461PubMedGoogle Scholar
  7. Calvert RC, Shabbir M, Thompson CS, Mikhailidis DP, Morgan RJ, Burnstock G (2004) Immunocytochemical and pharmacological characterisation of P2-purinoceptor-mediated cell growth and death in PC-3 hormone refractory prostate cancer cells. Anticancer Res 24:2853–2859PubMedGoogle Scholar
  8. Chiozzi P, Sanz JM, Ferrari D, Falzoni S, Aleotti A, Buell GN, Collo G, Di Virgilio F (1997) Spontaneous cell fusion in macrophage cultures expressing high levels of the P2Z/P2X7 receptor. J Cell Biol 138:697–706CrossRefPubMedGoogle Scholar
  9. Chueh SH, Kao LS (1993) Extracellular ATP stimulates calcium influx in neuroblastoma x glioma hybrid NG108–15 cells. J Neurochem 61:1782–1788PubMedGoogle Scholar
  10. Coutinho-Silva R, Persechini PM, Bisaggio RD, Perfettini JL, Neto AC, Kanellopoulos JM, Motta-Ly I, Dautry-Varsat A, Ojcius DM (1999) P2Z/P2X7 receptor-dependent apoptosis of dendritic cells. Am J Physiol 276:C1139–C1147Google Scholar
  11. Cowen DS, Berger M, Nuttle L, Dubyak GR (1991) Chronic treatment with P2-purinergic receptor agonists induces phenotypic modulation of the HL-60 and U937 human myelogenous leukemia cell lines. J Leukoc Biol 50:109–122PubMedGoogle Scholar
  12. Dreiling L, Hoffman S, Robinson WA (1996) Melanoma: epidemiology, pathogenesis, and new modes of treatment. Adv Intern Med 41:553–604PubMedGoogle Scholar
  13. Dubyak GR, De Young MB (1985) Intracellular Ca2+ mobilization activated by extracellular ATP in Ehrlich ascites tumor cells. J Biol Chem 260:10653–10661PubMedGoogle Scholar
  14. Ferrari D, La Sala A, Chiozzi P, Morelli A, Falzoni S, Girolomoni G, Idzko M, Dichmann S, Norgauer J, Di Virgilio F (2000) The P2 purinergic receptors of human dendritic cells: identification and coupling to cytokine release. FASEB J 14:2466–2476CrossRefPubMedGoogle Scholar
  15. Fredholm BB, IJzerman AP, Jacobson KA, Klotz KN, Linden J (2001) International Union of Pharmacology. XXV. Nomenclature and classification of adenosine receptors. Pharmacol Rev 53:527–552PubMedGoogle Scholar
  16. Gargett CE, Wiley JS (1997) The isoquinoline derivative KN-62 a potent antagonist of the P2Z-receptor of human lymphocytes. Br J Pharmacol 120:1483–1490Google Scholar
  17. Gartland A, Hipskind RA, Gallagher JA, Bowler WB (2001) Expression of a P2X7 receptor by a subpopulation of human osteoblasts. J Bone Miner Res 16:846–856PubMedGoogle Scholar
  18. Giard DJ, Aaronson SA, Todaro GJ, Arnstein P, Kersey JH, Dosik H, Parks WP (1973) In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst 51:1417–1423PubMedGoogle Scholar
  19. Gillies RJ, Didier N, Denton M (1986) Determination of cell number in monolayer cultures. Anal Biochem 159:109–113CrossRefPubMedGoogle Scholar
  20. Greig AV, Linge C, Healy V, Lim P, Clayton E, Rustin MH, McGrouther DA, Burnstock G (2003a) Expression of purinergic receptors in non-melanoma skin cancers and their functional roles in A431 cells. J Invest Dermatol 121:315–327CrossRefPubMedGoogle Scholar
  21. Greig AV, Linge C, Terenghi G, McGrouther DA, Burnstock G (2003b) Purinergic receptors are part of a functional signaling system for proliferation and differentiation of human epidermal keratinocytes. J Invest Dermatol 120:1007–1015CrossRefPubMedGoogle Scholar
  22. Haskell CM, Wong M, Williams A, Lee LY (1996) Phase I trial of extracellular adenosine 5′-triphosphate in patients with advanced cancer. Med Pediatr Oncol 27:165–173CrossRefPubMedGoogle Scholar
  23. Hoyle CHV, Burnstock G (1993) Postganglionic efferent transmission to the bladder and urethra. In: Maggi C (ed) The autonomic nervous system, vol 3. Nervous control of the urogenital system. Harwood Academic, Switzerland, pp 349–383Google Scholar
  24. Humphreys BD, Dubyak GR (1996) Induction of the P2z/P2X7 nucleotide receptor and associated phospholipase D activity by lipopolysaccharide and IFN-gamma in the human THP-1 monocytic cell line. J Immunol 157:5627–5637PubMedGoogle Scholar
  25. Humphreys BD, Rice J, Kertesy SB, Dubyak GR (2000) Stress-activated protein kinase/JNK activation and apoptotic induction by the macrophage P2X7 nucleotide receptor. J Biol Chem 275:26792–26798PubMedGoogle Scholar
  26. Janssens R, Boeynaems JM (2001) Effects of extracellular nucleotides and nucleosides on prostate carcinoma cells. Br J Pharmacol 132:536–546Google Scholar
  27. Khakh BS, Barnard EA, Burnstock G, Kennedy C, King BF, North RA, Séguéla P, Voigt M, Humphrey PPA (2000) P2X receptor-channels. In: Girdlestone D (ed) The IUPHAR compendium of receptor characterization and classification. IUPHAR Media Ltd, London, pp 290–305Google Scholar
  28. King BF, Burnstock G, Boyer JL, Boeynaems J-M, Weisman GA, Kennedy C, Jacobson KA, Humphries RG, Abbracchio MP, Gachet C, Miras-Portugal MT (2000) The P2Y receptors. In: Girdlestone D (ed) The IUPHAR compendium of receptor characterization and classification. IUPHAR Media Ltd, London, pp 306–320Google Scholar
  29. Lin WW, Chuang DM (1993) Extracellular ATP stimulates inositol phospholipid turnover and calcium influx in C6 glioma cells. Neurochem Res 18:681–687CrossRefPubMedGoogle Scholar
  30. Maaser K, Hopfner M, Kap H, Sutter AP, Barthel B, von Lampe B, Zeitz M, Scherubl H (2002) Extracellular nucleotides inhibit growth of human oesophageal cancer cells via P2Y(2)-receptors. Br J Cancer 86:636–644Google Scholar
  31. MacKenzie A, Wilson HL, Kiss-Toth E, Dower SK, North RA, Surprenant A (2001) Rapid secretion of interleukin-1beta by microvesicle shedding. Immunity 15:825–835CrossRefPubMedGoogle Scholar
  32. Mehta VB, Hart J, Wewers MD (2001) ATP-stimulated release of interleukin (IL)-1beta and IL-18 requires priming by lipopolysaccharide and is independent of caspase-1 cleavage. J Biol Chem 276:3820–3826CrossRefPubMedGoogle Scholar
  33. Merighi S, Varani K, Gessi S, Cattabriga E, Iannotta V, Ulouglu C, Leung E, Borea PA (2001) Pharmacological and biochemical characterization of adenosine receptors in the human malignant melanoma A375 cell line. Br J Pharmacol 134:1215–1226Google Scholar
  34. Merighi S, Mirandola P, Milani D, Varani K, Gessi S, Klotz KN, Leung E, Baraldi PG, Borea PA (2002) Adenosine receptors as mediators of both cell proliferation and cell death of cultured human melanoma cells. J Invest Dermatol 119:923–933CrossRefPubMedGoogle Scholar
  35. Nakamura E, Uezono Y, Narusawa K, Shibuya I, Oishi Y, Tanaka M, Yanagihara N, Nakamura T, Izumi F (2000) ATP activates DNA synthesis by acting on P2X receptors in human osteoblast-like MG-63 cells. Am J Physiol Cell Physiol 279:C510–C519Google Scholar
  36. Nicholson DW, Thornberry NA (1997) Caspases: killer proteases. Trends Biochem Sci 22:299–306PubMedGoogle Scholar
  37. Oglesby IB, Lachnit WG, Burnstock G, Ford AP (1999) Subunit specificity of polyclonal antisera to the carboxy terminal regions of P2X receptors, P2X1 through P2X7. Drug Dev Res 47:189–195CrossRefGoogle Scholar
  38. Osborne JE (2002) Skin cancer screening and surveillance. Br J Dermatol 146:745–754Google Scholar
  39. Per LK, Jon HA, Dissing S (2002) The human SH-SY5Y neuroblastoma cell-line expresses a functional P2X7 purinoceptor that modulates voltage-dependent Ca2+ channel function. J Neurochem 83:285–298CrossRefPubMedGoogle Scholar
  40. Popper LD, Batra S (1993) Calcium mobilization and cell proliferation activated by extracellular ATP in human ovarian tumour cells. Cell Calcium 14:209–218CrossRefPubMedGoogle Scholar
  41. Ralevic V, Burnstock G (1998) Receptors for purines and pyrimidines. Pharmacol Rev 50:413–492PubMedGoogle Scholar
  42. Rapaport E (1983) Treatment of human tumor cells with ADP or ATP yields arrest of growth in the S phase of the cell cycle. J Cell Physiol 114:279–283CrossRefPubMedGoogle Scholar
  43. Rapaport E, Fontaine J (1989) Generation of extracellular ATP in blood and its mediated inhibition of host weight loss in tumor-bearing mice. Biochem Pharmacol 38:4261–4266CrossRefPubMedGoogle Scholar
  44. Ryten M, Dunn PM, Neary JT, Burnstock G (2002) ATP regulates the differentiation of mammalian skeletal muscle by activation of a P2X5 receptor on satellite cells. J Cell Biol 158:345–355CrossRefPubMedGoogle Scholar
  45. Serrone L, Hersey P (1999) The chemoresistance of human malignant melanoma: an update. Melanoma Res 9:51–58PubMedGoogle Scholar
  46. Slater M, Scolyer RA, Gidley-Baird A, Thompson JF, Barden JA (2003) Increased expression of apoptotic markers in melanoma. Melanoma Res 13:137–145CrossRefPubMedGoogle Scholar
  47. Sneddon P, Burnstock G (1984) Inhibition of excitatory junction potentials in guinea-pig vas deferens by alpha, beta-methylene-ATP: further evidence for ATP and noradrenaline as cotransmitters. Eur J Pharmacol 100:85–90CrossRefPubMedGoogle Scholar
  48. Surprenant A, Rassendren F, Kawashima E, North RA, Buell G (1996) The cytolytic P2Z receptor for extracellular ATP identified as a P2X receptor (P2X7). Science 272:735–738PubMedGoogle Scholar
  49. Torres GE, Egan TM, Voigt MM (1999) Hetero-oligomeric assembly of P2X receptor subunits. Specificities exist with regard to possible partners. J Biol Chem 274:6653–6659CrossRefPubMedGoogle Scholar
  50. Wiley JS, Gargett CE, Zhang W, Snook MB, Jamieson GP (1998) Partial agonists and antagonists reveal a second permeability state of human lymphocyte P2Z/P2X7 channel. Am J Physiol 275:C1224–C1231Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Nicholas White
    • 1
  • Peter E. M. Butler
    • 2
  • Geoffrey Burnstock
    • 1
    Email author
  1. 1.Autonomic Neuroscience CentreRoyal Free and University College Medical SchoolLondonUK
  2. 2.Department of Plastic and Reconstructive SurgeryRoyal Free HospitalLondonUK

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