Figure 7. UCN-01 increases dl922-947 activity in vitro and in vivo.

(A) A2780CP and IGROV1 cells were infected with dl922-947. UCN-01 was added 6 hpi. Cell survival was assessed 120 hpi. (B) A2780CP cells were harvested 72 hours after dl922-947 (MOI 7.5) infection and treatment with UCN-01. Titers of intracellular and extracellular adenovirus were assessed by TCID₅₀. (C) A2780CP and IGROV1 cells were infected with dl922-947 or Ad5 WT. PF477736 (360 nM for A2780CP, 120 nM for IGROV1) was added 6 hpi. Cell survival was assessed 120 hpi. (D) dl922-947 (10¹⁰ particles) was injected into subcutaneous IGROV1 xenografts on the flank of athymic nu/nu female mice. 24 hpi, mice received a single i.p. dose of UCN-01 (7.5 mg/kg) or vehicle (20% icodextrin). Mice were killed 24 hours thereafter. Expression of E1A was assessed by immunohistochemistry (top). RNA was extracted from snap frozen tumors, and E1A expression was assessed by qRT-PCR (bottom). Expression was normalized to that of 18S RNA and plotted relative to mice that received dl922-947 and vehicle. Original magnification, ×40. **P < 0.01. (E) 5 × 10⁶ IGROV1-luciferase cells were injected i.p. into athymic nu/nu female mice. On day 9, mice were randomly allocated to receive i.p. dl922-947 or control adenovirus (5 × 10⁹ particles/day on days 9, 10, 15, and 16), with or without i.p. UCN-01 (5 mg/kg on days 11, 12, 17, and 18). Tumor burden was assessed using bioluminescence imaging. Bars represent SEM; n = 5/group. *P < 0.05, for 1-tailed comparison of dl922-947/UCN-01 versus both adenovirus GFP/UCN-01 and dl922-947/vehicle.