FIG. 7.

A constitutively activated form of AKT2, but not treatment with IGF-1, rescues human cancer cells from FTI-277-induced apoptosis. (A) OVCAR-3 cells were transfected with pcDNA₃ or constitutively active AKT2 (Myr-HA-AKT2), and two stable clones were established. Western blot analyses with an anti-HA antibody revealed expression of Myr-AKT2 in these two clonal cell lines but not in cells transfected with pcDNA₃ vector alone. (B and C) DNA fragmentation and TUNEL assay. After treatment with FTI-277, the DNA ladder and apoptotic cells were not observed in Myr-AKT2-transfected cells (lane 4 of panel B and middle column of panel C). However, FTI-277-induced apoptosis was detected in pcDNA₃-transfected OVCAR-3 cells cultured in medium containing either IGF-1 (50 ng/ml; lane 5 of panel B and rightmost column of panel C) or 5% FCS (lane 3 of panel B and leftmost column of panel C). Lane 1, φX174 replicative-form DNA/HaeIII markers; lanes 2 and 3, parental OVCAR-3 cells treated with vehicle (DMSO) and FTI-277, respectively.