Abstract
Glioblastoma is the most aggressive form of primary brain cancer, with a median survival of 12–15 months. The P2X receptor 7 (P2X7R) is upregulated in glioblastoma and is associated with increased tumor cell proliferation. The cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) is also upregulated in glioblastoma and has been shown to have both pro- and anti-tumor functions. This study investigates the potential mechanism linking P2X7R and GM-CSF in the U251 glioblastoma cell line and the therapeutic potential of P2X7R antagonism in this setting. P2X7R protein and mRNA was demonstrated to be expressed in the U251 cell line as assessed by immunocytochemistry and qPCR. Its channel function was intact as demonstrated by live cell confocal imaging using a calcium indicator Fluo-4 AM. Inhibition of P2X7R using antagonist AZ10606120, decreased both GM-CSF mRNA (P < 0.05) and protein (P < 0.01) measured by qPCR and ELISA respectively. Neutralization of GM-CSF with an anti-GM-CSF antibody did not alter U251 cell proliferation, however, P2X7R antagonism with AZ10606120 significantly reduced U251 glioblastoma cell numbers (P < 0.01). This study describes a novel link between P2X7R activity and GM-CSF expression in a human glioblastoma cell line and highlights the potential therapeutic benefit of P2X7R inhibition with AZ10606120 in glioblastoma.
Highlights
Abbreviations Analysis of Variance (ANOVA) Analysis of variance adenosine triphosphate (ATP) Adenosine triphosphate BzATP 3′-O-(4-Benzoyl)benzoyl ATP central nervous system (CNS) Central nervous system colony-stimulating factor 2 (CSF2) Colony-stimulating factor 2 DAPI 4′,6-Diamidino-2-phenylindole Dulbecco’s modified Eagle’s Medium (DMEM) Dulbecco’s modified eagles medium ELISA Enzyme linked immunosorbent assay FITC Fluorescein isothiocyanate GAPDH Glyceraldehyde 3-phosphate dehydrogenase GFAP Glial fibrillary acidic protein
The results of this study identified expression of both P2X7R and GM-CSFRα in the U-251 MG human glioblastoma cell line (U251) glioblastoma cell line and established that the P2X7R ion channel is functional in this setting
This study describes, for the first time, a link between P2X7R and granulocyte-macrophage colony-stimulating factor (GM-CSF) and the effect of P2X7R antagonism on cell proliferation in human glioblastoma cell line U251
Summary
Abbreviations ANOVA Analysis of variance ATP Adenosine triphosphate BzATP 3′-O-(4-Benzoyl)benzoyl ATP CNS Central nervous system CSF2 Colony-stimulating factor 2 DAPI 4′,6-Diamidino-2-phenylindole DMEM Dulbecco’s modified eagles medium ELISA Enzyme linked immunosorbent assay FITC Fluorescein isothiocyanate GAPDH Glyceraldehyde 3-phosphate dehydrogenase GFAP Glial fibrillary acidic protein. Gliomas are the most common primary malignancies of the central nervous system (CNS), representing 81% of malignant brain tumors in a dults[1,2] They are derived from glial cells, with an overall incidence between 3 and 6 per 100,000 p ersons[3,4,5]. The current standard therapy for glioblastoma includes surgical resection of the tumor, followed by radiotherapy and adjuvant chemotherapy using temozolomide (TMZ)[5,8]. Neuroinflammation plays a key role in the pathogenesis of glioblastoma It involves the activation and recruitment of immune cells such as microglia, monocytes and lymphocytes that can release a variety of bioactive factors that influence the tumor microenvironment. Neuroinflammatory mediators are of key interest in understanding the link between neuroinflammation and glioblastoma propagation[14]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
