P2X7 receptor activation induces cell death and CD23 shedding in human RPMI 8226 multiple myeloma cells

Biochimica Et Biophysica Acta
Andrew W FarrellRonald Sluyter

Abstract

The extracellular ATP-gated cation channel, P2X7 receptor, has an emerging role in neoplasia, however progress in the field is limited by a lack of malignant cell lines expressing this receptor. Immunofluorescence labelling and a fixed-time ATP-induced ethidium+ uptake assay were used to screen a panel of human malignant cell lines for the presence of functional P2X7. The presence of P2X7 was confirmed by RT-PCR, immunoblotting and pharmacological approaches. ATP-induced cell death was measured by colourimetric tetrazolium-based and cytofluorometric assays. ATP-induced CD23 shedding was measured by immunofluorescence labelling and ELISA. RPMI 8226 multiple myeloma cells expressed P2X7 mRNA and protein, as well as P2X1, P2X4 and P2X5 mRNA. ATP induced ethidium+ uptake into these cells with an EC₅₀ of ~116 μM, and this uptake was reduced in the presence of extracellular Ca²+ and Mg²+. The P2X7 agonist 2'- and 3'-0(4-benzoylbenzoyl) ATP, but not UTP, induced ethidium+ uptake. ATP-induced ethidium+ uptake was impaired by the P2X7 antagonists, KN-62 and A-438079. ATP induced death and CD23 shedding in RPMI 8226 cells, and both processes were impaired by P2X7 antagonists. The metalloprotease antagonists, BB-94 and GM6001, impaired AT...Continue Reading

Citations

Jun 5, 2012·Purinergic Signalling·Safina GadeockRonald Sluyter
Jun 26, 2013·Purinergic Signalling·Geoffrey Burnstock, Francesco Di Virgilio
Aug 15, 2013·Purinergic Signalling·Geoffrey BurnstockIsabel R Orriss
Jun 9, 2012·Cardiovascular Research·Alexander W LohmanBrant E Isakson
Oct 29, 2013·Purinergic Signalling·Wolfgang FischerMichael Schaefer
Sep 13, 2012·Journal of Osteoporosis·Elena AdinolfiAnna Lisa Giuliani
Feb 23, 2013·Biochemical and Biophysical Research Communications·Aleta PupovacRonald Sluyter
Jan 28, 2014·British Journal of Pharmacology·I AzimiG R Monteith
Dec 3, 2013·Veterinary Immunology and Immunopathology·Mari SpildrejordeRonald Sluyter
Jan 17, 2015·Journal of Molecular Endocrinology·Ankita Agrawal, Alison Gartland
Aug 27, 2014·Immunology and Cell Biology·Aleta PupovacRonald Sluyter
May 18, 2016·Cellular and Molecular Life Sciences : CMLS·Aleta Pupovac, Ronald Sluyter
Jul 5, 2013·PloS One·Carolyn GlassArchibald Perkins
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Jun 28, 2019·Journal of Hematology & Oncology·Shengchao XuChunfang Zhang
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Apr 29, 2021·Signal Transduction and Targeted Therapy·Zhao HuangYong Tang
Sep 29, 2021·Cell Communication and Signaling : CCS·Tingting LiZhiyong Zeng

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