P2Y 1 nucleotide receptor silencing and its effect on glioma C6 calcium signaling

• Autorzy: Wypych D. , Pomorski P.
• Języki publikacji: EN

Abstrakty

EN

In our earlier studies of the signaling cross-talk between nucleotide receptors in an in vitro glioma model (C6 cell line) under prolonged serum deprivation conditions, a growth arrest of the cells and expression shift from P2Y1 to P2Y12 receptors was found. The aim of the present work was to test if siRNA silencing of P2Y1 receptor changes P2Y12 expression similarly as following the serum deprivation and which physiological downstream pathways it affects. Here we demonstrate for the first time the efficiency of siRNA technology in silencing P2Y nucleotide receptors in glioma C6 cell line. Moreover, P2Y12 proved to be insensitive to the P2Y1 receptor silencing. The effect of the P2Y1 silencing on calcium signaling was less pronounced then the extent of the protein change itself, exactly as was the case for the serum starvation experiments. Phosphorylation of ERK and Akt kinases were studied as the downstream effect of P2Y1-evoked signaling and similar effects as in the case of serum deprivation were found for ERK, and even stronger ones for Akt phosphorylation.

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2012
• Tom: 59
• Numer: 4
• Opis fizyczny: p.711-717,fig.,ref.

Twórcy

autor

Wypych D.

• Laboratory of Molecular Basis of Cell Motility, Department of Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland

autor

Pomorski P.

• Laboratory of Molecular Basis of Cell Motility, Department of Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland

Bibliografia

• Abbracchio MP, Burnstock G (1998) Purinergic signalling: pathophysiological roles. Jpn J Pharmacol 78: 113-145. 
• Auer RN, Del Maestro RF, Anderson R (1981) A simple and reproducible experimental in vivo glioma model. Can J Neurol Sci 8: 325-331. 
• Bagchi S, Liao Z, Gonzalez FA, Chorna NE, Seye CI, Weisman GA, Erb L (2005) The P2Y2 nucleotide receptor interacts with alphav integrins to activate Go and induce cell migration. J Biol Chem 280: 39050-39057. 
• Baranska J, Chaban V, Czarny M, Sabala P (1995) Changes in Ca2+ concentration in phorbol ester and thapsigargin treated glioma C6 cells. The role of protein kinase C in regulation of Ca2+ entry. Cell calcium 17: 207-215. 
• Benda P, Lightbody J, Sato G, Levine L, Sweet W (1968) Differentiated rat glial cell strain in tissue culture. Science 161: 370-371. 
• Braganhol E, Huppes D, Bernardi A, Wink MR, Lenz G, Battastini AM (2009) A comparative study of ectonucleotidase and P2 receptor mRNA profiles in C6 cell line cultures and C6 ex vivo glioma model. Cell Tissue Res 335: 331-340. 
• Brismar T (1995) Physiology of transformed glial cells. Glia 15: 231-243. 
• Burnstock G (1976) Purinergic receptors. J Theor Biol 62: 491-503. 
• Burnstock G (2007) Physiology and pathophysiology of purinergic neurotransmission. Physiol Rev 87: 659-797. 
• Crooke A, Mediero A, Guzman-Aranguez A, Pintor J (2009) Silencing of P2Y2 receptor delays Ap4A-corneal re-epithelialization process. Mol Vis 15: 1169-1178. 
• Czajkowski R, Banachewicz W, Ilnytska O, Drobot LB, Baranska J (2004) Differential effects of P2Y1 and P2Y12 nucleotide receptors on ERK1/ERK2 and phosphatidylinositol 3-kinase signalling and cell proliferation in serum-deprived and nonstarved glioma C6 cells. Br J Pharmacol 141: 497-507. 
• De Vuyst E, Decrock E, Cabooter L, Dubyak GR, Naus CC, Evans WH, Leybaert L (2006) Intracellular calcium changes trigger connexin 32 hemichannel opening. EMBO J 25: 34-44. 
• Deli T, Varga N, Adam A, Kenessey I, Raso E, Puskas LG, Tovari J, Fodor J, Feher M, Szigeti GP, Csernoch L, Timar J (2007) Functional genomics of calcium channels in human melanoma cells. Int J Cancer 121: 55-65. 
• Edelstein A, Amodaj N, Hoover K, Vale R, Stuurman N (2010) Computer control of microscopes using microManager. Current Protocols in Molecular Biology. Frederick M. Ausubel FM et al. eds. Chapter 14: Unit14 20.
• Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260: 3440-3450. 
• Harden TK, Sesma JI, Fricks IP, Lazarowski ER (2010) Signalling and pharmacological properties of the P2Y receptor. Acta Physiol (Oxford, England) 199: 149-160. 
• Hardy AR, Jones ML, Mundell SJ, Poole AW (2004) Reciprocal cross-talk between P2Y1 and P2Y12 receptors at the level of calcium signaling in human platelets. Blood 104: 1745-1752. 
• Jacobs VL, Valdes PA, Hickey WF, De Leo JA (2011) Current review of in vivo GBM rodent models: emphasis on the CNS-1 tumour model. ASN Neuro 3: e00063. 
• Krzeminski P, Suplat D, Czajkowski R, Pomorski P, Baranska J (2007) Expression and functional characterization of P2Y1 and P2Y12 nucleotide receptors in long-term serum-deprived glioma C6 cells. FEBS J 274: 1970-1982. 
• Krzeminski P, Pomorski P, Baranska J (2008) The P2Y14 receptor activity in glioma C6 cells. Eur J Pharmacol 594: 49-54. 
• Liao Z, Seye CI, Weisman GA, Erb L (2007) The P2Y2 nucleotide receptor requires interaction with alpha v integrins to access and activate G12. J Cell Sci 120: 1654-1662. 
• Martin-Gil A, de Lara MJ, Crooke A, Santano C, Peral A, Pintor J (2012) Silencing of P2Y(2) receptors reduces intraocular pressure in New Zealand rabbits. Br J Pharmacol 165: 1163-1172. 
• North RA (2002) Molecular physiology of P2X receptors. Physiol Rev 82: 1013-1067. 
• O'Brien KA, Gartner TK, Hay N, Du X (2012) ADP-stimulated activation of Akt during integrin outside-in signaling promotes platelet spreading by inhibiting glycogen synthase kinase-3beta. Arterioscler Thromb Vasc Biol 32: 2232-2240. 
• Ohsawa K, Irino Y, Sanagi T, Nakamura Y, Suzuki E, Inoue K, Kohsaka S (2010) P2Y12 receptor-mediated integrin-beta1 activation regulates microglial process extension induced by ATP. Glia 58: 790-801. 
• Rasmussen H, Jensen P, Lake W, Goodman DB (1976) Calcium ion as second messenger. Clin Endocrinol 5 (Suppl) 11S-27S. 
• Sak K, Illes P (2005) Neuronal and glial cell lines as model systems for studying P2Y receptor pharmacology. Neurochem Int 47: 401-412. 
• Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9: 671-675. 
• Suplat-Wypych D, Dygas A, Baranska J (2010) 2', 3'-O-(4-benzoylbenzoyl)-ATP-mediated calcium signaling in rat glioma C6 cells: role of the P2Y(2) nucleotide receptor. Purinergic Signal 6: 317-325. 
• Suplat D, Krzeminski P, Pomorski P, Baranska J (2007) P2Y(1) and P2Y(12) receptor cross-talk in calcium signalling: Evidence from nonstarved and long-term serum-deprived glioma C6 cells. Purinergic Signal 3: 221-230. 
• Suzuki N, Nakamura S, Mano H, Kozasa T (2003) Galpha 12 activates Rho GTPase through tyrosine-phosphorylated leukemia-associated RhoGEF. Proc Natl Acad Sci USA 100: 733-738. 
• Van Kolen K, Slegers H (2004) P2Y12 receptor stimulation inhibits beta-adrenergic receptor-induced differentiationby reversing the cyclic AMP-dependent inhibition of protein kinase B. J Neurochem 89: 442-453. 
• Wei W, Ryu JK, Choi HB, McLarnon JG (2008) Expression and function of the P2X(7) receptor in rat C6 glioma cells. Cancer Lett 260: 79-87. 
• Wypych D, Baranska J (2013) Cross-talk in nucleotide signaling in glioma C6 cells. Adv Exp Med Biol 986: 31-59. 

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