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Notably, co administration of PHA 739358 with FTI resulted in a striking increase in the sub G1 compartment, To determine the ability of PHA 739358 to augment the efficacy of drugs currently in use in a clinical setting for therapy of Ph ALL, we treated Pt2 cells with 2. 5 nM or 5. 0 nM vincristine alone or together with 1 uM PHA 739358 for 3 days. As demon strated in Additional Maraviroc CCR5 阻害剤 file 1, Figure S1A, exposure of Pt2 to 2. 5 nM or 5. 0 nM vincristine alone decreased cell viability to 80 and 50%, respectively. The combined treatment with PHA 739358 and vincristine further significantly reduced cell viability and cell numbers. A combination of dasatinib with PHA 739358 in wild type Bcr Abl UCSF02 had a similar effect, The growth inhibitory effect of PHA 739358 on human ALL cells was further confirmed using a colony formation assay.<br><br> As shown in Additional file 2, Figure S2, 10 nM PHA 739358 led MK-2206 to about 55% and 25% re duction of colony numbers in Pt2 and UCSF02 cells, re spectively, compared with the controls. PHA 739358 at a concentration of 25 nM almost completely inhibited the colony formation of both Pt2 and UCSF02 cells. Combined treatment of PHA 739358 with FTI, vincristine or dasatinib completely inhibited the growth of Pt2 and UCSF02 as assessed by colony formation assay. Therefore, we confirmed that a significant portion of the effect of PHA 739358 on human ALL cells was due to its growth inhibitory effect. In vivo efficacy of PHA 739358 on Bcr Abl cells with T315I mutation To examine the efficacy of PHA 739358 in vivo, Pt2 cells with the T315I mutation were transplanted into NSG mice via tail vein injection.<br><br> After mice developed leukemia, we evaluated the inhibitory effects of PHA 739358 on the phosphorylation levels of tyrosine, histone mtorc2 阻害剤 H3 and Crkl 2 hours after drug administration. As shown in Figure 5, there was a significant down regulation of the levels of total phosphotyrosine, of phospho Crkl and of phospho histone H3 by Western blot, both in bone mar row and spleen of mice transplanted with leukemia cells, indicating that it was able to inhibit both Bcr Abl and Aurora B activities in vivo. We also measured the effect of PHA 739358 on the out come of leukemia. Seven days after transplantation of Pt2 ALL cells into NSG mice, we administered three cycles of 30 mg kg PHA 739358 treatment. One cycle consisted of daily injections for 7 days, followed by a 7 day break.<br><br> We monitored the percentage of leukemia cells in the periph eral blood by flow cytometry. Figure 6A, B shows that, in comparison with vehicle treated mice, PHA 739358 trea ted mice showed significantly decreased amounts of leukemia cells in the peripheral blood on day 32, day 46 and day 59 after transplantation. However, peripheral blood still contained around 5% of leukemia cells even after two cycles of PHA 739358 treatment at day 32. When the administration of PHA 739358 was terminated on day 42, leukemia cells started to proliferate again in the treatment group. Figure 6B demonstrates that from day 46 to day 59, the per centage of leukemia cells in the PHA 739358 treated group increased from about 10% to 40%, compared to the control group in which an increase from 55% to 70% was measured.