title: P2Y1 nucleotide receptor silencing and its effect on glioma C6 calcium signaling.
abstract: In our earlier studies of the signaling cross-talk between nucleotide receptors in an in vitro glioma model (C6 cell line) under prolonged serum deprivation conditions, a growth arrest of the cells and expression shift from P2Y(1) to P2Y(12) receptors was found. The aim of the present work was to test if siRNA silencing of P2Y(1) receptor changes P2Y(12) expression similarly as following the serum deprivation and which physiological downstream pathways it affects. Here we demonstrate for the first time the efficiency of siRNA technology in silencing P2Y nucleotide receptors in glioma C6 cell line. Moreover, P2Y(12) proved to be insensitive to the P2Y(1) receptor silencing. The effect of the P2Y(1) silencing on calcium signaling was less pronounced then the extent of the protein change itself, exactly as was the case for the serum starvation experiments. Phosphorylation of ERK and Akt kinases were studied as the downstream effect of P2Y(1)-evoked signaling and similar effects as in the case of serum deprivation were found for ERK, and even stronger ones for Akt phosphorylation.
authors.affiliation: Laboratory of Molecular Basis of Cell Motility, Department of Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland.
authors.forename: Dorota
authors.initials: D
authors.lastname: Wypych
authors.forename: Pawel
authors.initials: P
authors.lastname: Pomorski
mesh_terms: D000818:Animals; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology; D020013:Calcium Signaling / Q000235:genetics; D045744:Cell Line, Tumor; D016895:Culture Media, Serum-Free; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D005910:Glioma / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D051058:Oncogene Protein v-akt / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D058472:Receptors, Purinergic P2Y1 / Q000235:genetics / Q000378:metabolism; D058925:Receptors, Purinergic P2Y12 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D016895:Culture Media, Serum-Free; D008565:Membrane Proteins; D058472:Receptors, Purinergic P2Y1; D058925:Receptors, Purinergic P2Y12; D051058:Oncogene Protein v-akt