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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Adenosine Induced Apoptosis in BHK Cells via P1 Receptors and Equilibrative Nucleoside Transporters

  • Sun, Wentian (Department of Biochemistry, Faculty of Medicine, National University of Singapore) ;
  • Khoo, Hoon Eng (Department of Biochemistry, Faculty of Medicine, National University of Singapore) ;
  • Tan, Chee Hong (Department of Biochemistry, Faculty of Medicine, National University of Singapore)
  • Published : 2005.05.31
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Abstract

Adenosine, as a ubiquitous metabolite, mediates many physiological functions via activation of plasma membrane receptors. Mechanisms of most of its physiological roles have been studied extensively, but research on adenosine-induced apoptosis (AIA) has only started recently. In this study we demonstrate that adenosine dose-dependently triggered apoptosis of cultured baby hamster kidney (BHK) cells. Adenosine-induced apoptotic cell death was characterized by DNA laddering, changes in nuclear chromatin morphology and phosphatidylserine staining. Apoptosis was also quantified by flow cytometry. Results suggest the involvement of adenosine $A_1$ and $A_3$ receptors as well as equilibrative nucleoside transporters in apoptosis induced by adenosine. These results indicate a receptor-transporter co-signaling mechanism in AIA in BHK cells. The involvement of $A_1$ and $A_3$ receptors also implies a possible apoptotic pathway mediated by G protein-coupled receptors.

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References

  1. Abbracchio, M. P., Ceruti, S., Barbieri, D., Franceschi, C., Malorni, W., Biondo, L., Burnstock, G. and Cattabeni, F. (1995) A novel action for adenosine: apoptosis of astroglial cells in rat brain primary cultures. Biochem. Biophys. Res. Commun. 213, 908-915 https://doi.org/10.1006/bbrc.1995.2215
  2. Abbracchio, M. P. and Williams, M. (2001a) Purinergic and pyrimidinergic signalling I. Springer-Verlag, Berlin, Germany
  3. Abbracchio, M.P., and Williams, M. (2001b) Purinergic and pyrimidinergic signalling II. Springer-Verlag, Berlin, Germany
  4. Barbieri, D., Abbracchio, M. P., Salvioli, S., Monti, D., Cossarizza, A., Ceruti, S., Brambilla, R., Cattabeni, F., Jacobson, K. A. and Franceschi, C. (1998) Apoptosis by 2- chloro-2'-deoxy-adenosine and 2-chloro-adenosine in human peripheral blood mononuclear cells. Neurochem. Int. 32, 493- 504 https://doi.org/10.1016/S0197-0186(97)00129-0
  5. Cass, C. E., Young, J. D. and Baldwin, S. A. (1998) Recent advances in the molecular biology of nucleoside transporters of mammalian cells. Biochem. Cell Biol. 76, 761-770 https://doi.org/10.1139/bcb-76-5-761
  6. Ceruti, S., Barbieri, D., Veronese, E., Cattabeni, F., Cossarizza, A., Giammarioli, A. M., Malorni, W., Franceschi, C. and Abbracchio, M. P. (1997) Different pathways of apoptosis revealed by 2-chloro-adenosine and deoxy-D-ribose in mammalian astroglial cells. J. Neurosci. Res. 47, 372-383 https://doi.org/10.1002/(SICI)1097-4547(19970215)47:4<372::AID-JNR2>3.0.CO;2-B
  7. Chow, S. C., Kass, G. E. and Orrenius, S. (1997) Purines and their roles in apoptosis. Neuropharmacology 36, 1149-1156 https://doi.org/10.1016/S0028-3908(97)00123-8
  8. Di Iorio, P., Kleywegt, S., Ciccarelli, R., Traversa, U., Andrew, C. M., Crocker, C. E., Werstiuk, E. S. and Rathbon, M. P. (2002) Mechanisms of apoptosis induced by purine nucleosides in astrocytes. Glia 38, 179-190 https://doi.org/10.1002/glia.10055
  9. Griffith, D. A. and Jarvis, S. M. (1996) Nucleoside and nucleobase transport systems of mammalian cells. Biochim. Biophys. Acta 1286, 153-181 https://doi.org/10.1016/S0304-4157(96)00008-1
  10. Homburg, C. H., de Haas, M., von dem Borne, A. E., Verhoeven, A. J., Reutelingsperger, C. P. and Roos, D. (1995) Human neutrophils lose their surface Fc gamma RIII and acquire Annexin V binding sites during apoptosis in vitro. Blood 85, 532-540
  11. Kohno, Y., Sei, Y., Koshiba, M., Kim, H. O., Jacobson, K. A. and Shainberg, A. (1998) Induction of apoptosis in HL-60 human promyelocytic leukemia cells by adenosine A(3) receptor agonists. Exp. Cell Res. 243, 383-397 https://doi.org/10.1006/excr.1998.4134
  12. Koshiba, M., Kosaka, H., Nakazawa, T., Hayashi, N., Saura, R., Kitamura, N. and Kumagai, S. (2002) 2-Chloroadenosine but not adenosine induces apoptosis in rheumatoid fibroblasts independently of cell surface adenosine receptor signaling. Br. J. Pharmacol. 135, 1477-1486 https://doi.org/10.1038/sj.bjp.0704612
  13. Liu, Y. and Kitsis, R. N. (1996) Induction of DNA synthesis and apoptosis in cardiac myocytes by E1A oncoprotein. J. Cell Biol. 133, 325-334 https://doi.org/10.1083/jcb.133.2.325
  14. Lodish, H., Berk, A., Zipursky, S. L., Matsudaira, P., Baltimore, D. and Darnell, J. E. (2000) Cell-to-cell signalling: hormones and receptors; in Molecular Cell Biology, Tenny, S. (ed.), pp. 852-854, W. H. Freeman and Co., New York, USA
  15. Martin, S. J., Reutelingsperger, C. P., McGahon, A. J., Rader, J. A., van Schie, R. C., LaFace, D. M. and Green, D. R. (1995) Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: inhibition by overexpression of Bcl-2 and Abl. J. Exp. Med. 182, 1545-1556 https://doi.org/10.1084/jem.182.5.1545
  16. Merighi, S., Mirandola, P., Milani, D., Varani, K., Gessi, S., Klotz, K. N., Leung, E., Baraldi, P. G. and Borea, P. A. (2002) Adenosine receptors as mediators of both cell proliferation and cell death of cultured human melanoma cells. J. Invest. Dermatol. 119, 923-933 https://doi.org/10.1046/j.1523-1747.2002.00111.x
  17. Palmer, T. M. and Stiles, G. L. (1995) Adenosine receptors. Neuropharmacology 34, 683-694 https://doi.org/10.1016/0028-3908(95)00044-7
  18. Parkinson, F. E. and Fredholm, B. B. (1991) Effects of propentofylline on adenosine A1 and A2 receptors and nitrobenzylthioinosine-sensitive nucleoside transporters: quantitative autoradiographic analysis. Eur. J. Pharmacol. 202, 361-366 https://doi.org/10.1016/0014-2999(91)90279-Y
  19. Parkinson, F. E., Paterson, A. R., Young, J. D. and Cass, C. E. (1993) Inhibitory effects of propentofylline on [3H]adenosine influx. A study of three nucleoside transport systems. Biochem. Pharmacol. 46, 891-896 https://doi.org/10.1016/0006-2952(93)90499-M
  20. Peyot, M. L., Gadeau, A. P., Dandre, F., Belloc, I., Dupuch, F. and Desgranges, C. (2000) Extracellular adenosine induces apoptosis of human arterial smooth muscle cells via A(2b)- purinoceptor. Circ. Res. 86, 76-85 https://doi.org/10.1161/01.RES.86.1.76
  21. Ralevic, V. and Burnstock, G. (1998) Receptors for purines and pyrimidines. Pharmacol. Rev. 50, 413-492
  22. Rimon, G., Bazenet, C. E., Philpott, K. L. and Rubin, L. L. (1997) Increased surface phosphatidylserine is an early marker of neuronal apoptosis. J. Neurosci. Res. 48, 563-570 https://doi.org/10.1002/(SICI)1097-4547(19970615)48:6<563::AID-JNR9>3.0.CO;2-8
  23. Rounds, S., Yee, W. L., Dawicki, D. D., Harrington, E., Parks, N. and Cutaia, M. V. (1998) Mechanism of extracellular ATP- and adenosine-induced apoptosis of cultured pulmonary artery endothelial cells. Am. J. Physiol. 275, 379-388
  24. Schrier, S. M., van Tilburg, E. W., van der Meulen, H., Ijzerman, A. P., Mulder, G. J. and Nagelkerke, J. F. (2001) Extracellular adenosine-induced apoptosis in mouse neuroblastoma cells: studies on involvement of adenosine receptors and adenosine uptake. Biochem. Pharmacol. 61, 417-425 https://doi.org/10.1016/S0006-2952(00)00573-6
  25. Schrier, S. M., Florea, B. I., Mulder, G. J., Nagelkerke, J. F. and Ijzerman, A. P. (2002) Apoptosis induced by extracellular ATP in the mouse neuroblastoma cell line N1E-115: studies on involvement of P2 receptors and adenosine. Biochem. Pharmacol. 63, 1119-1126 https://doi.org/10.1016/S0006-2952(01)00939-X
  26. Shneyvays, V., Nawrath, H., Jacobson, K. A. and Shainberg, A. (1998) Induction of apoptosis in cardiac myocytes by an A3 adenosine receptor agonist. Exp. Cell Res. 243, 383-397 https://doi.org/10.1006/excr.1998.4134
  27. Szondy, Z. (1994) Adenosine stimulates DNA fragmentation in human thymocytes by $Ca^{2+}$-mediated mechanisms. Biochem. J. 304, 877-885
  28. Szondy, Z. (1995) The 2-chlorodeoxyadenosine-induced cell death signalling pathway in human thymocytes is different from that induced by 2-chloroadenosine. Biochem. J. 311, 585-588
  29. Tanaka, Y., Yoshihara, K., Tsuyuki, M. and Kamiya, T. (1994) Apoptosis induced by adenosine in human leukemia HL-60 cells. Exp. Cell Res. 213, 242-252 https://doi.org/10.1006/excr.1994.1196
  30. Wakade, T. D., Palmer, K. C., McCauley, R., Przywara, D. A. and Wakade, A. R. (1995) Adenosine-induced apoptosis in chick embryonic sympathetic neurons: a new physiological role for adenosine. J. Physiol. 488, 123-138 https://doi.org/10.1113/jphysiol.1995.sp020951
  31. Walker, B. M., Rocchini, C., Boone, R. H., Ip, S. and Jacobson, M. A. (1997) Adenosine A2a receptor activation delays apoptosis in human neutrophils. J. Immunol. 158, 2926-2931
  32. Yao, Y., Sei, Y., Abbracchio, M. P., Jiang, J. L., Kim, Y. C. and Jacobson, K. A. (1997) Adenosine A3 receptor agonists protect HL-60 and U-937 cells from apoptosis induced by A3 antagonists, Biochem. Biophys. Res. Commun. 232, 317-322 https://doi.org/10.1006/bbrc.1997.6290

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