L Peng1, C J Bradley, J S Wiley. 1. Department of Laboratory Medicine, First Hospital, West China University of Medical Sciences, Chengdu 610041, China.
Abstract
OBJECTIVE: To investigate the role of purinergic P2Z receptors for apoptosis of human leukemic lymphocytes mediated by extracellular adenosine triphosphate (ATP). METHODS: A total of 13 B-chronic lymphocytic leukemia (CLL) patients were studied. Exposure of leukemic lymphocytes with (n = 8) or without (n = 5) P2Z receptors to ATP, benzoylbenzoic-ATP (BzATP), 2-methylthio-ATP (2MeSATP), adenosine-5' [gamma-thio] triphosphate (ATP-gamma S), and other nucleosides for 8 h in vitro. Apoptosis was detected by electron microscopy (EM), agarose gel electrophoresis, and the quantitative assay-TdT assay. RESULTS: Apoptosis was detected only in leukemic lymphocytes with P2Z receptors. Using a quantitative assay, ATP-induced DNA strand breaks were found to occur specifically with BzATP, ATP and 2MeSATP, but not for analogue ATP-gamma S nor other nucleosides. Meanwhile, ATP-induced DNA fragmentation was fully blocked by pretreatment with oxidized ATP (OxATP), a compound recently shown to block P2Z receptors. Also, it is shown that the Ca2+/calmodulin complex plays a role in the regulation of the apoptosis induced by ATP on CLL cells, because an antagonist of this complex, 1-[N, O-bis (5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62) was found to inhibit the ATP-induced apoptosis. Furthermore, choline, an inhibitor of phospholipase D (PLD), is first shown to partially inhibit ATP-induced apoptosis. CONCLUSION: These data indicate that P2Z receptors on lymphocytes play an important role in the apoptosis induced by ATP in vitro.
OBJECTIVE: To investigate the role of purinergic P2Z receptors for apoptosis of humanleukemic lymphocytes mediated by extracellular adenosine triphosphate (ATP). METHODS: A total of 13 B-chronic lymphocytic leukemia (CLL) patients were studied. Exposure of leukemic lymphocytes with (n = 8) or without (n = 5) P2Z receptors to ATP, benzoylbenzoic-ATP (BzATP), 2-methylthio-ATP (2MeSATP), adenosine-5' [gamma-thio] triphosphate (ATP-gamma S), and other nucleosides for 8 h in vitro. Apoptosis was detected by electron microscopy (EM), agarose gel electrophoresis, and the quantitative assay-TdT assay. RESULTS: Apoptosis was detected only in leukemic lymphocytes with P2Z receptors. Using a quantitative assay, ATP-induced DNA strand breaks were found to occur specifically with BzATP, ATP and 2MeSATP, but not for analogue ATP-gamma S nor other nucleosides. Meanwhile, ATP-induced DNA fragmentation was fully blocked by pretreatment with oxidized ATP (OxATP), a compound recently shown to block P2Z receptors. Also, it is shown that the Ca2+/calmodulin complex plays a role in the regulation of the apoptosis induced by ATP on CLL cells, because an antagonist of this complex, 1-[N, O-bis (5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62) was found to inhibit the ATP-induced apoptosis. Furthermore, choline, an inhibitor of phospholipase D (PLD), is first shown to partially inhibit ATP-induced apoptosis. CONCLUSION: These data indicate that P2Z receptors on lymphocytes play an important role in the apoptosis induced by ATP in vitro.
Authors: Birgitte R Kornum; Minae Kawashima; Juliette Faraco; Ling Lin; Thomas J Rico; Stephanie Hesselson; Robert C Axtell; Hedwich Kuipers; Karin Weiner; Alexandra Hamacher; Matthias U Kassack; Fang Han; Stine Knudsen; Jing Li; Xiaosong Dong; Juliane Winkelmann; Giuseppe Plazzi; Sona Nevsimalova; Seung-Chul Hong; Yutaka Honda; Makoto Honda; Birgit Högl; Thanh G N Ton; Jacques Montplaisir; Patrice Bourgin; David Kemlink; Yu-Shu Huang; Simon Warby; Mali Einen; Jasmin L Eshragh; Taku Miyagawa; Alex Desautels; Elisabeth Ruppert; Per Egil Hesla; Francesca Poli; Fabio Pizza; Birgit Frauscher; Jong-Hyun Jeong; Sung-Pil Lee; Kingman P Strohl; William T Longstreth; Mark Kvale; Marie Dobrovolna; Maurice M Ohayon; Gerald T Nepom; H-Erich Wichmann; Guy A Rouleau; Christian Gieger; Douglas F Levinson; Pablo V Gejman; Thomas Meitinger; Paul Peppard; Terry Young; Poul Jennum; Lawrence Steinman; Katsushi Tokunaga; Pui-Yan Kwok; Neil Risch; Joachim Hallmayer; Emmanuel Mignot Journal: Nat Genet Date: 2010-12-19 Impact factor: 38.330