Effect of ectopic overexpression of survivin on VG-1 cells.

Ectopic overexpression of survivin rescues VG-1 cells from apoptosis induced by AG490. (A) Survivin-transfected and control vector–transfected VG-1 cells were treated with AG490 for 24 hours, and whole cell lysates (25 μg per lane) were subjected to SDS-PAGE. Western blotting demonstrates that survivin expression is unchanged in survivin-transfected but is reduced in control vector–transfected cells after AG490 treatment. (B) Cells were incubated with the indicated concentrations of AG490 for 24 hours, including a final 6-hour pulse with [³H]-thymidine. The results represent the mean percentage (± SD) of radioactivity as compared with cultures in 0.1% DMSO alone. Shown is a representative experiment of 3 performed. (C) Effects of enforced survivin expression on AG490-induced programmed cell death. After incubation with AG490 or 0.1% DMSO for 48 hours, cells were stained with annexin V–FITC and analyzed for apoptosis by flow cytometry. AG490 induced marked increase of the annexin V⁺ population in vector-transfected control cells but not in survivin-transfected cells. Data shown are representative of 3 independent experiments. The percentage of annexin V–FITC-binding cells are shown.