Effect of Cdc25C protein expression on the proliferation of PCa cells.

<p>(A,B) LNCaP C-33 and (C) PC-3 cells were plated at a density of 1.2×10⁴ or 1×10⁴ cells/cm² for 72 hr and then transfected with Cdc25C cDNA (A) or shRNA (B,C) plasmids. Control cells were transfected with the respective vector alone. (A) Cdc25C cDNA transfected C-33 cells were cultured in regular medium and the cell number was then counted after 2 days. (B) Cdc25C shRNA-transfected C-33 cells were steroid starved for 48 hr in SR medium, and then treated with or without 10 nM DHT for 2 days. (C) Cdc25C shRNA transfected PC-3 cells were cultured in regular medium for 2 days, and the cell number was counted. The ratio of cell proliferation was calculated by normalizing the experimental cell number to that of control cells transfected with vector alone, respectively. Total cell lysate proteins were analyzed for Cdc25A, Cdc25B, Cdc25C, Cyclin B1 and/or Cyclin D1 proteins. β-actin was used as a loading control. The ratios of Cdc25C, Cyclin B1 and CyclinD1 protein levels to β-actin were calculated after semi-quantification by densitometric analyses on films with different exposure time periods. *<i>p</i><0.05, n = 2×3; Bar, standard deviation.