Also kinases associated with Src pathway were highly active. In addition, retinoic acid recep tor pathway and peroxisome proliferator acti vated receptor activation pathway were found. The top 5 of activated pathways was identical sellekchem in cell lines and primary cultures. Results of the analysis leav ing out all cell cycle related kinases, which might be artificially upregulated due to cell culturing, and results of analysis Inhibitors,Modulators,Libraries after starvation of the cell lines are shown in table 3. Verification of kinome profiling Western blotting showed that all myxoid liposarcoma samples expressed comparable amounts of total Src and Inhibitors,Modulators,Libraries NF kappaB p65. Phosphorylation of Src was present in all sam ples confirming activation of Src pathway.

Likewise, western blotting showed the presence of ck2a1 and phosphorylated NF kappaB p65 in all sam ples, confirming the results of the IPA analysis that kinases associated with NF kappaB pathway are active in myxoid liposarcoma cells. In vitro Inhibitors,Modulators,Libraries targeting of kinases associated with Src and NF kappaB pathways by dasatinib and TBB WST 1 analysis of GIST882 showed a profound decrease in cell viability of up to 80% relative to the DMSO control at even low dosages of Src inhibitor dasatinib. The decrease in cell viability of myxoid liposarcoma cells treated with dasatinib was rather mild as WST 1 analysis of all four cell cultures and 1 out of 2 cell lines showed a maximum decrease in cell viability of 40% at higher doses. Cell line 1765 92 did not respond to dasatinib. In contrast, myx oid liposarcoma cells showed a decline of more than 50% in viability after treatment with casein kinase 2 inhibitor TBB in two out of four cultures and in both cell lines.

This effect was also observed in Jurkat cells as described. There was no relation between the response rate and type of fusion gene. For combination experiments, the two cell lines and the two most sensitive myxoid liposar coma primary Inhibitors,Modulators,Libraries cultures were treated with both dasatinib and TBB. Combined administration of both drugs led to a dramatic decrease in cell viability and showed an enhanced effect, for instance L1357 cells show 80% viability at maximum dasatinib dose, whereas viability was only 5% at lower concentration of dasatinib at IC 50 for TBB. Dasatinib inhibits phosphorylation of Src but does not cause apoptosis To investigate the effect of dasatinib on Src signalling, a good responsive myxoid liposarcoma cell culture was treated with 50, 200 and 500 nM of dasatinib for 6 hours.

Whereas levels of total Src did not visibly decrease upon dasatinib treatment, a decrease in phosphorylated Src was Inhibitors,Modulators,Libraries found. At a dose of 200 nM dasatinib p Src staining the lower band faded and at 500 nM both bands disappeared. Interestingly, a similar decrease in p Src was also observed at 200 nM dasatinib when post treated CHIR99021 with TBB.