Conclusions Roscovitine Dovitinib mechanism has shown to be able to significantly modify the DDR response. Even considering the many genes that are potentially involved, the putative role of CDK2 in multiple DDR pathways along with the downregula tion of cyclin A1, may further explain the effective inhi bition of a broad range of DNA repair mechanisms by Roscovitine. In particular since NHEJ is considered the major pathway for the repair of gIR induced DNA DSBs in human cells, we believe our data support further investigation on the therapeutic advantages of combina tion therapy with Roscovitine and Radiotherapy.

Methods Cell Culture and Serum Starvation The following Inhibitors,Modulators,Libraries solid cancer human cell lines were pur chased from and authenticated by American Type Cul ture Collection and cultured at 37 C in a humidified atmosphere of 5% CO2 in air, within the appropriate medium according to supplier recommendations supplemented with 10% heat inactivated fetal bovine serum and 100U of Penicillin and 100 ug/ml of Streptomycin NSCLC cell lines A549 and H23, breast cancer cell lines MCF 7 and MDA MB 231, prostate cancer cell lines LNCAP and DU145, and the adenovirus transformed human embryonic kidney epithelial cells HEK293FT. Cells were regularly Inhibitors,Modulators,Libraries sub cultured according to ATCC recommen dations with a 0. 25% trypsin EDTA solution. To obtain synchronous populations Inhibitors,Modulators,Libraries of cells, confluent plates of A549 cells were incubated in media supplemented with 0. 1% heat inactivated fetal bovine serum for 96 hours. Cells were then sub cultured into serum con taining medium and time points were taken every four hours.

Drugs, irradiations and treatments Doxorubicin was obtained from BioMol International. Lyopholized drug was re sus pended into a 1 1 mixture of dimethyl sulfoxide and MilliQ fil tered H2O to a concentration of 4. 31 mM, aliquoted for use and stored Inhibitors,Modulators,Libraries at 20 C. Ros covitine was obtained from Signa Gen Laboratories. Lyophilized drug was re suspended into DMSO to a concentration of 14. 1 mM, aliquoted and stored at 20 C until use. Inhibitors,Modulators,Libraries Fresh dilutions from the stock solutions were prepared for each treatment. Taxol was obtained from USB Corporation. Lyophilized drug was re suspended into DMSO to a concentration of 5. 86 mM, aliquoted and stored at 20 C until use. MG 132 was obtained from Sigma. Lyophilized drug was re suspended into DMSO to a concentration of 10 mg/ml, aliquoted and stored at 20 C until use.

Irradiations were performed in an AECL Gamma Cell 40, Cs 137 irradiator at a dose rate of 1 Gy/minute for respective doses. In treatments throughout this article the control samples refer to cells treated with an equal selleck DAPT secretase concentration of DMSO as in the highest drug concentration used per experiment. Western Blot Analysis and SDS PAGE Equal amounts of whole cell lysates were resolved by SDS PAGE and transferred to a nitrocellu lose membrane by wet electrophoretic transfer.