Figure 6

Responses of tissue slice cultures (TSCs) to androgen ablation. Culture of benign and prostate cancer (PCa) TSCs in the absence of androgen for 5 days resulted in heterogeneous reduction of cellular viability and glandular integrity according to MTS assays (a) and hematoxylin and eosin (H&E) staining (b, arrows point to degenerating glands), respectively. Luminal degeneration was also evident upon immunohistochemistry (IHC) evaluation of benign and PCa TSCs cultured for 5 days in the absence (ethanol control) of R1881 (c). Benign and PCa TSCs were verified by α-methylacyl coenzyme A racemase (AMACR)/p63/cytokeratin 5 (CK5) staining (top row). Androgen ablation reduced the number of actively proliferating cells (Ki67 positive, black arrows) and increased the number of cells undergoing apoptosis (cleaved caspase 3 (CC3) positive, red arrows) in both benign and PCa TSCs. Prostate-specific antigen (PSA) expression was reduced in both benign and PCa TSCs upon 5 days of androgen ablation (bottom row). (d) The percentages of CC3- and Ki67-positive nuclei were quantified from six random × 40 fields from each of three tissue slices per condition (average of 90 nuclei per field). *P<0.05, **P<0.01. Experiments and representative images were from patient specimens number 18, 20, and 21.