Viability of SH-SY5Y cells is associated with purinergic P2 receptor expression alterations.
To investigate the role of metabotrophic purinergic P2Y receptors in neuroblastoma cell survival, expression of P2 receptors by normal mouse (C57BL/6) brain and human neuroblastoma SH-SY5Y cells was investigated by Western blot and real time PCR studies. Viability of SH-SY5Y cells treated with purinergic receptor antagonists suramin and pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS) was evaluated by MTT assay and flow cytometry. In the brain samples of C57BL/6 mice, expressions of P2Y4 and P2X7 were significantly reduced, whereas that of P2Y1 was significantly elevated in an age-dependent manner. SH-SY5Y cell viability was significantly reduced and necrotic cell rates were mildly increased by 400 μM suramin and 100 μM PPADS treatment. Antagonist treatment downregulated P2Y1, P2Y2 and P2Y4 and upregulated P2Y6, P2Y12 and P2X7 mRNA levels in SH-SY5Y cells on the 24th hour. These alterations were abolished for all P2 receptors except P2Y1 in the 48th hour. P2Y receptors are expressed by both normal mouse brain and human neuroblastoma cells. Purinergic receptor antagonism interferes with neuroblastoma viability through elevation of necrotic cell death and modulation of P2 receptor expression. P2Y receptors might thus be useful targets for future anti-tumor treatment trials.
Department of Neuroscience, Aziz Sancar Institute of Experimental Medicine, Istanbul University , Istanbul , Turkey.
Arda
A
Örçen
Department of Neuroscience, Aziz Sancar Institute of Experimental Medicine, Istanbul University , Istanbul , Turkey.
Vuslat
V
Yilmaz
Department of Neuroscience, Aziz Sancar Institute of Experimental Medicine, Istanbul University , Istanbul , Turkey.
Murat
M
Giriş
Department of Neuroscience, Aziz Sancar Institute of Experimental Medicine, Istanbul University , Istanbul , Turkey.
Uğur
U
Akcan
Department of Neuroscience, Aziz Sancar Institute of Experimental Medicine, Istanbul University , Istanbul , Turkey.
Erdem
E
Tüzün
Department of Immunology, Aziz Sancar Institute of Experimental Medicine, Istanbul University , Istanbul , Turkey.
Gaye
G
Erten
D000367:Age Factors; D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D008297:Male; D008810:Mice, Inbred C57BL; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D058919:Purinergic P2 Receptor Antagonists / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013498:Suramin / Q000494:pharmacology
D020033:Protein Isoforms; D058919:Purinergic P2 Receptor Antagonists; D018048:Receptors, Purinergic P2; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; D011732:Pyridoxal Phosphate; D013498:Suramin
MTT-viability; Neuroblastoma; SH-SY5Y; purinergic receptor