If AZD1152 or other AURKB inhibitors could be demonstrated to raise the therapeutic index for androgen resistant prostate cancer, this would have a significant clinical effect. Cell Culture and Reagents PC 3 and DU145 cells were cultured in RPMI 1640 medium containing 10 % fetal bovine serum and 1% penicillin/streptomycin. All cells were incubated at 37 C in 95-pound air/5% CO2. AZD1152 was obtained from AstraZeneca. Western Immunoblotting natural products research Cells were treated with various concentrations of AZD1152. These were collected at different times and then washed with ice-cold PBS twice before the addition of lysis buffer including protease inhibitor cocktail and phosphatase inhibitor cocktail I. Protein concentration was quantified by the Bio Rad technique. Equal amounts of protein were separated by 12 and loaded into each well. 5% or 15,000-25,000 SDS PAGE gel, followed by transfer onto PVDF membranes. Membranes were blocked with five hundred non-fat dry milk in PBST for 1 h at room temperature. The blots were then incubated with anti phosphohistone H3, anti Aurora B, and anti Actin for 1 h at 4 C. Western blots were developed utilizing the Retroperitoneal lymph node dissection chemiluminescence detection system in line with the producers protocol and autoradiography. Mobile Cycle Analysis Cells were then treated with various doses of AZD1152 for 48 h and seeded in 10 cm2 meals 24 h before treatment. The cells were then obtained by trypsinization, fixed with 70-300mm ethanol, and stored over night at C. Cells were then collected by centrifugation, resuspended in 1 ml of PBS with 100 ul of 200 g/ml DNase free RNase A, and incubated at 37 C for 30 min. Propidium iodide was then added, and the cells were incubated HDAC6 inhibitor at room temperature for 5 min. The number of cells in each stage of the cell cycle was determined and calculated as a share of the total cell population. 8 Gy/min using a 137Cs irradiator. After irradiation, the medium was modified and cells were incubated at 37 C for 8 days. Cells were then set for 30 min with 70-300 methanol and stained for 30 min with hands down the methylene blue in water. After staining, colonies were counted by eye with a cut-off of 50 viable cells. The surviving fraction was calculated as / plating effectiveness, where PE was understood to be /. Cells were then washed twice in PBS and positioned on cover slips in ice cold wells. Then 2 ml of ice-cold Triton X 100 solution was added. After 15 min, the cover slips were washed 3 times in PBS.