3), both considered a hallmark of apoptosis [41] and [42]. Interestingly, the type of death signal generated in the two cell lines seems to differ and be cell type-dependent. In this Cobimetinib clinical trial respect, we show that PCP treatment of MIA PaCa-2 cells leads to activation of both the extrinsic and intrinsic caspase-mediated apoptotic pathways as indicated by the cleavage of caspase-8 and caspase-9, respectively, and the dose-dependent decreased level of cytochrome c in the mitochondria (Fig. 5). In the case of Panc-1 cells, induction

of cell death is mediated solely by the death receptor-mediated caspase pathway as indicated by the cleavage of caspase-8 and lack of significant decrease in the levels of mitochondrial cytochrome c as compared to control cells. Loss of mitochondrial www.selleckchem.com/products/Everolimus(RAD001).html membrane potential is believed to occur during activation of death pathways and accompanied by cytochrome c release. As shown in Fig. 5, both cell lines lose their membrane

potential during C11 or PCP-induced apoptosis as indicated by the remarkable decrease in the JC-1 red fluorescence signal. Surprisingly, it appeared that decreased ΔΨm did not correlate with cytochrome c release in Panc-1 cells suggesting that these two events occur independently from each other. In support of these data, Johnson et al. [43] proposed that the mitochondria contribute to the activation of death pathways

at various levels and that release of cytochrome c and mitochondria depolarization are separate and independent events depending on where the Avelestat (AZD9668) contribution of the mitochondria in the death pathway resides. The analysis of intracellular signalling pathways that have been shown to be de-regulated in pancreatic cancer supporting growth and conferring chemoresistance, suggest that the cytotoxic properties of PCP are not solely confined to the inhibition of CK2 but also to alteration of other intracellular signalling molecules. In this respect, phosphorylation of JNK was found up-regulated. JNK is part of a family of protein kinases activated in response to a wide range of cellular stresses [44]. Hence, increased phosphorylation observed following C11 or PCP treatment might represent a stress response accompanying activation of the apoptotic cell death signalling as previously postulated [45]. Unexpectedly, the anti-proliferative response of PCP correlated with increased phosphorylation of AKT S473 and T308 and a mild effect on AKT protein expression levels in Mia PaCa-2 cells (Fig. 6b). At a first glance these results may appear contradictory as the PI3K/AKT signalling pathway has been linked to cell growth and survival and, thus, one would expect that this signalling cascade would remain unaltered or be suppressed during induction of cell death.