New Pyrazolopyrimidine Inhibitors of Protein Kinase D as Potent Anticancer Agents for Prostate Cancer Cells
Figure 6
1-NA-PP1 inhibited PC3 cell proliferation, survival, and arrested cells in G2/M.
A.1-NA-PP1 blocked PC3 cell proliferation. PC3 cells were plated in triplicates in 24-well plates. Cells were allowed to attach overnight. A cell count at day 1 was made, and then either a vehicle (DMSO) or 1-NA-PP1 at 10 µM was added. Cells were counted daily for a total of 5 days. Fresh media and inhibitor were added every 2 days. The means of triplicate determinations were plotted over time. The experiment was repeated twice and results from one representative experiment are shown. B. 1-NA-PP1 induced cell death in PC3 cells. PC3 cells were seeded into 96-well plates (3000 cells/well) and were then incubated in media containing 0.3–100 µM inhibitors for 72 h. MTT solution was added to each well and incubated for 4 h. Optical density was read at 570 nm to determine cell viability. The IC50 was determined as the mean of two independent experiments for each compound. C. 1-NA-PP1 caused G2/M phase cell cycle arrest. PC3 cells were treated with either vehicle (DMSO), or 10 µM 1-NA-PP1 for 48 h. Cell cycle distribution was determined by flow cytometry after propidium iodide labeling of fixed cells. Statistical significance was determined by unpaired t-test and is indicated. **, p<0.01; ***, p<0.001.