Fig. 3: MDM2 inhibitors produced synergistic growth inhibitory effects and changed cell cycle progression with Ad-delE1B on wild-type p53 cells.

A Wild-type p53 or B mutated p53 cells were infected with various vp of Ad-delE1B for 2 days, and then treated with nutlin-3a (MSTO-211H: 0.5 µM, NCI-H226, EHMES-1, and JMN-1B: 1 µM) or RG7112 (0.15 µM) for further 2 days. The cell viabilities were measured with the WST agent. Relative viability was calculated based on untreated cells. Averages and SE bars are shown (n = 3). CI values were plotted at respective Fa points. CI < 1, CI = 1 and CI > 1 indicate synergistic, additive, and antagonistic actions, respectively. C Cells were infected with Ad-delE1B (MSTO-211H and NCI-H226: 1.5 × 10³ vp/cell, EHMES-1 and JMN-1B: 3 × 10³ vp/cell) for 2 days, and then treated with nutlin-3a (MSTO-211H and NCI-H226: 5 µM, EHMES-1, and JMN-1B: 15 µM) or RG7112 (MSTO-211H and NCI-H226: 3 µM, EHMES-1 and JMN-1B: 10 µM). Live cells numbers were then counted on 24 and 48 h later. ns not significant, *p < 0.001 (GraphPad Prism 6, La Jolla). D Cells were infected with Ad-delE1B or Ad-LacZ (3 × 10³ vp/cell) for 2 days, and then treated with nutlin-3a (5 µM) or RG7112 (5 µM). Cell cycle distribution was determined with flow cytometry. Representative histogram of MSTO-211H cells at 36 h after treatment with the inhibitor and that of NCI-H226 at 48 h. E Percentages of annexin V-positive cells. Cells were treated as above and annexin V-positive cells were determined with flow cytometry in MSTO-211H cells at 36 h after the inhibitor treatment and in NCI-H226 at 48 h. Data are presented as averages with SE bars (n = 3). *p < 0.01.