5C and D, there was an increase over time in the number of cells in mitotic catastrophe after individual treatments with radiation and AZD6244 at least 96 hours. In cells, the ASA404 DMXAA combination therapy had detected a significant increase in the percentage of cells containing a mitotic catastrophe were at 72 hours after treatment in cell lines of both the A549 and MiaPaCa2. This result was accompanied by an increase in the proportion of cells with more than 4n DNA content by flow cytometry. Proliferation of cells that are more than 4n DNA was within 24 hours after irradiation in both cell lines were treated with vehicle or AZD6244. In addition, the cells with DNA w were During 4n significantly A549 and MiaPaCa2 cells were treated with AZD6244 compared with the Tr hunter alone 96 hours the treated obtained after irradiation Ht.
These data suggest that AZD6244-mediated radiosensitization by the failure of the recovery is then conveyed after irradiation caused no increase in the continuous cell mitotic catastrophe. To determine whether the improvement of the radiation sensitivity of tumor cells was measured in vitro in a tumor model in vivo assay Tumorwachstumsverz Extension with A549 and MiaPaCa2 cells subcutaneously in the thigh of the raised Nacktm be translated Mice was. Mice that sc xenografts were randomized into four groups: vehicle AZD6244, AZD6244 alone IR alone and administered by oral gavage 4 hours before IR. The treatment was the day of randomization. The growth rate for A549 and MiaPaCa2 tumors are exposed to each treatment in Figure 6 A and B are each presented.
For each group was time to get from 172 to 1500 mm3 mm3 grow calculated with the B ends of tumors in each of Mice in each group. For A549 xenograft model, the time required for tumors of cro To 172 to 1500 mm 3 for 24.8 days for Mice with vehicle 1.0 to 40.0 1.7 days for Mice treated with AZD6244 increased Hten value. Treatment with radiation alone increased ht The necessary time to 1500 to 35.6 mm3 1.5 days to reach. However, in mice M That again If the combination of U AZD6244 IR time for tumors to grow to 1,500 mm 3 increased Hte to 61.4 1.9 days. The absolute growth delay in your transportation from 15.2 to 50 mg / kg AZD6244 alone, and 10.8 for irradiation alone, was the delay Gerung of tumor growth by AZD6244 treatment-induced IR 36.6.
Thus, the growth delay Gerung after a combined treatment more than the sum of the growth of individual delay Wrestled treatments caused. Gain for one dose Rkungsfaktor comparing the tumor response in M AZD6244 mice with and without irradiation were plated Siege to calculate tumor growth standard, the contribution of AZD6244 in a delay Gerung of tumor growth induced by the combined treatment. Normalized delay Gerung of tumor growth was defined as the time in days for tumors of 172-1500 mm 3 defined growth in M Mice, the combined modality t less time in days for tumors of 172 to grow to 1500 mm 3 at M Mice with AZD6244 alone were treated. The dose Gain Rkungsfaktor by the normalized Tumorwachstumsverz Storage in M Mice treated with AZD6244 IR M by growth retardation Mice that were treated with radiation only absolute is 3.38 to 50 mg / kg AZD6244. A Hnliches experiment was performed MiaPaCa2 xenografts. The growth rates for MiaPaCa2 tumors exposed to ch