Androgen/AR signaling leads to DNA damage in DPCs.

<p>AR was overexpressed in DPCs by transfected with pcDNA3-hAR and cultured in the presence of DHT of 0.1 μM or ethanol for 2 hours. (A) γ-H2AX foci was analyzed by immunofluorescence assays. DHT increased γ-H2AX foci in the nuclei of DPCs; AR overexpression reinforced the effect of DHT. Nuclei were counterstained with DAPI. Scale bar = 10 μm. Foci in cell nuclei were visualized on a microscope using a 40X objective. (B) The mean number of foci per cell was calculated from 40 cells randomly selected for each group. AR overexpression enhanced the statistical significance of DHT effects. Values are means ± SDs from three independent experiments. Asterisk indicates <i>P</i><0.05. (C) A representative immunoblot of cell lysates from DPCs after treatment. The numbers indicate γ-H2AX/H2AX and AR/GAPDH ratios. GAPDH was used as an internal standard. (D) Quantitative densitometry of the γ-H2AX/H2AX protein expression was performed by using ImageJ software. Values are means ± SDs from six independent experiments.