Figure 3
AdLCY displayed potent cytolytic activity against hypoxic bladder cancer cells while sparing normal cells. (a) Schematic diagram showing the genome organization of three recombinant adenoviruses, Ad5WS1, Ad9OC and AdLCY, as well as wild-type adenovirus type 5 (Ad5wt). Two point mutations were made at bases 2253 (C-T) and 2262 (G-T) of the E1B-55 kD gene (nucleotide 2019–3059) of Ad5 to generate premature translation stop codons, resulting in Ad5WS1, an E1B-55 kD-deleted adenovirus driven by the E1A promoter. ITR, inverted terminal repeat; ψ, packaging signal. Numbers indicate nucleotide positions of Ad5. The E1A promoter was replaced with 9 × ORE-CMVmini and 6 × HRE-9 × ORE -CMVmini promoters to generate Ad9OC and AdLCY, respectively. (b) Virus-induced CPE. Bladder cancer (TCC-SUP and MBT-2) and normal epithelial (SV-HUC-1 and NMuMG) cells were infected with increasing MOIs of AdLCY or Ad9OC, or were mock-infected under normoxia (N) or hypoxia (H) for 48 h. Cells were then cultured in normoxic conditions for 5 days, and the CPE was visualized by crystal violet staining.
