Potentiation of Ad-p53–induced apoptosis.

Bcl-2-ASO potentiates Ad-p53–induced apoptosis in ARH-77, U266 (A), and ARP-1 (B) cells. Cells were pretreated with or without 5 μg/mL Bcl-2-ASO for 3 days. Cultures were further incubated with Bcl-2-ASO plus low-dose (1000 vp/cell) or high-dose (2000 vp/cell) Ad-p53 for an additional 2 days. Cultures treated for 2 days with high- or low-dose Ad-p53 alone were also performed. Apoptosis was determined by annexin V. Bcl-2-ASO was replenished every other day. Bars are ± SD of at least 3 experiments. (C) Western immunoblotting for detection of p53 in Ad-p53–transduced cells. Cells were cultured as in panels A-B, and aliquots were taken for protein extraction and Western immunoblotting as described in the legend to Figure 1. Lane 1 indicates control, untreated cells; lanes 2 and 3, cells transduced for 1 day with low- and high-dose Ad-p53, respectively; lane 4, cells treated with Bcl-2-ASO for 5 days; lanes 5 and 6, cells pretreated for 3 days with Bcl-2-ASO followed by 2 days of transduction with low- and high-dose Ad-p53, respectively.