The presence of OTX2 (orthodenticle homeobox 2), a

homeobox protein acting as a transcription factor during brain development, seems to be necessary for ATRA-induced mortality of tumor cells. In accordance, enhanced OTX2 protein levels have been observed in the sensitive selleck products D283-Med cells, whereas the relatively resistant DAOY cells do not express OTX2 [41]. The combinatorial treatment with 5-aza-dC revealed no further effect in the ATRA-sensitive D283-Med cells but led to a significant increase of metabolic activity in DAOY cells compared to 5-aza-dC alone. The simultaneous treatment of the ATRA-resistant MEB-Med8a cells showed no 5-aza-dC-dependent effect on the ATRA responder status GDC-0449 in vitro (Figure 3d). In contrast, Fu et al. reported a 5-aza-dC-induced hypomethylation of the hypermethylated CRABP-II (cellular retinoic acid-binding protein) gene promoter in ATRA-resistant MB cells leading to the expression of the afore-silenced gene. This affects the ATRA transport into the nucleus and lead to an ATRA-mediated cellular response in these MB cells [47]. However, the lack of BMN 673 cell line an ATRA response in MEB-Med8a after combined treatment

with 5-aza-dC indicates that hypermethylation of the CRABP-II promoter is not responsible for ATRA resistance in this MB cell line. As shown in Figure 2e, resveratrol (> 10 μM) led to a significant concentration-dependent reduction of metabolic activity in all three examined cell lines, possibly by inhibition of STAT3 (signal transducer and activator of transcription 3) expression

and activity, which results in irreversible cell cycle arrest or apoptosis [44]. The IC 30 values of 15 μM (D283-Med, DAOY) and 40 μM (MEB-Med8a) are within the concentrations of 40 μM, maximal achievable in blood serum after intravenous injection [42]. The combined administration of resveratrol and 5-aza-dC showed a significant synergistic inhibition of 18% (MEB-Med8a), 41% (D283-Med) and 54% (DAOY) on metabolic activity versus 5-aza-dC alone (Figure 3e). The sensitive response of the TP53-mutated DAOY cell line might indicate a speculative role of resveratrol in the therapy of highly aggressive and therapy-resistant TP53-mutated MB Cediranib (AZD2171) tumors. Numerous studies, regarding the outcome of TP53-mutated MBs, which represents about 10% of all MBs, showed a 5-year event-free survival of 0% [43–47]. Interestingly, resveratrol has been shown to induce apoptosis p53-dependently and also p53-independently [48, 49]. Combinatorial effects of 5-aza-dC and resveratrol on clonogenicity and DSB repair Our investigations on metabolic activity revealed that 5-aza-dC combined with resveratrol achieve the highest antitumor response compared to the other tested drugs. To assess long-time effects, we determined the reproductive cell survival by clonogenic assay after combined 5-aza-dC and resveratrol treatment. 5-Aza-dC alone resulted in a decrease of surviving clonogenic cells exhibiting surviving fractions (SF) between 0.0014 (DAOY, D283-Med8) and 0.