FIG. 5.
Loss of RASSF2 protects prostate cancer cells from TRAIL-induced apoptosis. (A and B) PC-3 cells stably transfected with a RASSF2 shRNA construct or a control vector were seeded at 1 × 105cells/well in six-well plates and treated with 100 ng/ml TRAIL, and cell death was estimated 72 h later by trypan blue exclusion. Bars show the means of triplicate experiments, and standard deviations are indicated. *, P < 0.05 compared to control cells. (C) Similar experiments were then performed to measure apoptosis. These included a second RASSF2 shRNA cell line (# 2). Cells were seeded at 3 × 103 cells/well in 96-well plates, and caspase-3 and -7 activities were measured after TRAIL treatment. Caspase activity is expressed relative to results in untreated cells. The bars show means of duplicate experiments. with standard deviations shown. *, significantly different (P < 0.05) from cells transfected with a scrambled shRNA. (D) Western blot analysis of RASSF2 expression in PC-3 prostate cancer cells stably expressing a RASSF2 shRNA (#2) construct (#2) or scrambled shRNA. TFIIH was used as a loading control.