P2X7 receptor activation induces cell death and microparticle release in murine erythroleukemia cells

P Constantinescu; B Wang; K Kovacevic; I Jalilian; GJCGM Bosman; JS Wiley; R Sluyter

Journal article

P2X7 receptor activation induces cell death and microparticle release in murine erythroleukemia cells

P Constantinescu, B Wang, K Kovacevic, I Jalilian, GJCGM Bosman, JS Wiley, R Sluyter

Biochimica Et Biophysica Acta Biomembranes | Published : 2010

DOI: 10.1016/j.bbamem.2010.06.002

Abstract

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 EC50 of ~154μM. 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-12+ uptak..

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University of Melbourne Researchers

Patrick Constantinescu Author

Grants

Funding Acknowledgements

This work was supported by Cure Cancer Australia and the University of Wollongong (Wollongong, Australia). R.S. was a recipient of a European Travel Award from the Australian Academy of Science. We gratefully acknowledge Dr. Jason D. McArthur (University of Wollongong) and Ms. Yvonne Groenen-Dopp (Rijnstate Hospital, Arnhem, The Netherlands) for technical advice regarding RT-PCR and MP assays respectively, and Ms. Margaret Phillips (University of Wollongong) for technical assistance.