title: The metabotropic P2Y4 receptor participates in the commitment to differentiation and cell death of human neuroblastoma SH-SY5Y cells.
abstract: Extracellular nucleotides exert a variety of biological actions through different subtypes of P2 receptors. Here we characterized in the human neuroblastoma SH-SY5Y cells the simultaneous presence of various P2 receptors, belonging to the P2X ionotropic and P2Y metabotropic families. Western blot analysis detected the P2X1,2,4,5,6,7 and P2Y1,2,4,6, but not the P2X3 and P2Y12 receptors. We then investigated which biological effects were mediated by the P2Y4 subtype and its physiological pyrimidine agonist UTP. We found that neuronal differentiation of the SH-SY5Y cells with dibutiryl-cAMP increased the expression of the P2Y4 protein and that UTP itself was able to positively interfere with neuritogenesis. Moreover, transient transfection and activation of P2Y4 also facilitated neuritogenesis in SH-SY5Y cells, as detected by morphological phase contrast analysis and confocal examination of neurofilament proteins NFL. This was concurrent with increased transcription of immediate-early genes linked to differentiation such as cdk-5 and NeuroD6, and activity of AP-1 transcription family members such as c-fos, fos-B, and jun-D. Nevertheless, a prolonged activation of the P2Y4 receptor by UTP also induced cell death, both in naive, differentiated, and P2Y4-transfected SH-SY5Y cells, as measured by direct count of intact nuclei and cytofluorimetric analysis of damaged DNA. Taken together, our data indicate that the high expression and activation of the P2Y4 receptor participates in the neuronal differentiation and commitment to death of SH-SY5Y cells.
authors.affiliation: Fondazione Santa Lucia, Neurobiology Unit, Rome, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Cavaliere
authors.forename: Valeria
authors.initials: V
authors.lastname: Nestola
authors.forename: Susanna
authors.initials: S
authors.lastname: Amadio
authors.forename: Nadia
authors.initials: N
authors.lastname: D'Ambrosi
authors.forename: Daniela F
authors.initials: DF
authors.lastname: Angelini
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Sancesario
authors.forename: Giorgio
authors.initials: G
authors.lastname: Bernardi
authors.forename: Cinzia
authors.initials: C
authors.lastname: Volonté
mesh_terms: D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism*; D003994:Bucladesine / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D017781:Genes, Immediate-Early / Q000187:drug effects / Q000235:genetics; D006801:Humans; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000378:metabolism; D058910:Purinergic P2 Receptor Agonists; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000187:drug effects / Q000235:genetics; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology; D014544:Uridine Triphosphate / Q000378:metabolism / Q000494:pharmacology
chemical_list: D016900:Neurofilament Proteins; D058910:Purinergic P2 Receptor Agonists; D018048:Receptors, Purinergic P2; D014157:Transcription Factors; C044882:neurofilament protein L; C104749:purinoceptor P2Y4; D003994:Bucladesine; D014544:Uridine Triphosphate