P2Y1 nucleotide receptor silencing and its effect on glioma C6 calcium signaling
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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 P2Y1 to P2Y12 receptors was found. The aim of the present work was to test if siRNA silencing of P2Y1 receptor changes P2Y12 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, P2Y12 proved to be insensitive to the P2Y1 receptor silencing. The effect of the P2Y1 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 P2Y1-evoked signaling and similar effects as in the case of serum deprivation were found for ERK, and even stronger ones for Akt phosphorylation.
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