Document Detail


P2X7 receptor activation induces cell death and microparticle release in murine erythroleukemia cells.
MedLine Citation:
PMID:  20529664     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Extracellular ATP induces cation fluxes in and impairs the growth of murine erythroleukemia (MEL) cells in a manner characteristic of the purinergic P2X7 receptor, however the presence of P2X7 in these cells is unknown. This study investigated whether MEL cells express functional P2X7. RT-PCR, immunoblotting and immunofluorescence staining demonstrated the presence of P2X7 in MEL cells. Cytofluorometric measurements demonstrated that ATP induced ethidium+ uptake into MEL cells in a concentration-dependent fashion and with an EC(50) of approximately 154 microM. The most potent P2X7 agonist 2'- and 3'-0(4-benzoylbenzoyl) ATP, but not ADP or UTP, induced ethidium+ uptake. ATP-induced ethidium+ and YO-PRO-1(2+) uptake were impaired by the P2X7 antagonist, A-438079. A colourmetric assay demonstrated that ATP impaired MEL cell growth. A cytofluorometric assay showed that ATP induced MEL cell death and that this process was impaired by A-438079. Finally, cytofluorometric measurements of Annexin-V binding and bio-maleimide staining demonstrated that ATP could induce rapid phosphatidylserine exposure and microparticle release in MEL cells respectively, both of which were impaired by A-438079. These results demonstrate that MEL cells express functional P2X7, and indicate that activation of this receptor may be important in the death and release of microparticles from red blood cells in vivo.
Authors:
Patrick Constantinescu; Bin Wang; Kati Kovacevic; Iman Jalilian; Giel J C G M Bosman; James S Wiley; Ronald Sluyter
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-06-08
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1798     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-07-19     Completed Date:  2010-10-18     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1797-804     Citation Subset:  IM    
Copyright Information:
2010 Elsevier B.V. All rights reserved.
Affiliation:
School of Biological Sciences, University of Wollongong, Wollongong, Australia.
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MeSH Terms
Descriptor/Qualifier:
Adenosine Triphosphate / pharmacology
Animals
Apoptosis*
Cell Line, Tumor
Ethidium / metabolism
Leukemia, Erythroblastic, Acute / metabolism,  pathology*
Mice
Phosphatidylserines / physiology
Receptors, Purinergic P2 / physiology*
Chemical
Reg. No./Substance:
0/Phosphatidylserines; 0/Receptors, Purinergic P2; 0/purinergic P2X7 receptor; 3546-21-2/Ethidium; 56-65-5/Adenosine Triphosphate

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