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P2Y1 nucleotide receptor silencing and its effect on glioma C6 calcium signaling

100%
Wypych D., Pomorski P.
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2012
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vol. 59
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issue 4
711-717
EN
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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Cross-talk between the ATP and ADP nucleotide receptor signalling pathways in glioma C6 cells.

63%
Czajkowski R., Barańska J.
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2002
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vol. 49
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issue 4
877-889
EN
In this review we summarize the present status of our knowledge on the enzymes involved in the extracellular metabolism of nucleotides and the receptors involved in nucleotide signalling. We focus on the mechanism of the ATP and ADP signalling pathways in glioma C6, representative of the type of nonexcitable cells. In these cells, ATP acts on the P2Y2 receptor coupled to phospholipase C, whereas ADP on two distinct P2Y receptors: P2Y1 and P2Y12. The former is linked to phospholipase C and the latter is negatively coupled to adenylyl cyclase. The possible cross-talk between the ATP-, ADP- and adenosine-induced pathways, leading to simultaneous regulation of inositol 1,4,5-trisphosphate and cAMP mediated signalling, is discussed.
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