Figure 4. B02 synergizes with AZD1775 and radiation in vivo in an orthotopic mouse model of oral tongue cancer.

A, hematoxylin and eosin (H&E) and TUNEL-positive staining determined at day 5 in the 3D culture organoids made from HPV-positive HNSCC (MDA-HN-2C) cells following treatment with equivalent physiological dose of B02 and AZD1775 for 5 days as described in Methods. B, quantification of the positive TUNEL staining shown in A. C and D, tumor growth curves in orthotopic mouse model oral tongues bearing HNSCC (PCI13-G245D and UM-SCC-47) cells following treatment with B02 (50 mg/kg), AZD1775 (35 mg/kg) either alone or in combination as described in methods. E and F, Kaplan-Meier analysis for survival and relative body weight loss during the treatment in UM-SCC-47 tumor bearing mice. Each treatment group contains 9-10 mice. G, representative clonogenic survival images of HPV-negative HNSCC (HN31) cells pre-treated with B02 two hours before radiation (XRT) as indicated. H, percentage of surviving colonies normalized to untreated control. Combinations of B02 plus XRT caused more tumor cell death than XRT or B02 alone. I, tumor growth curves in orthotopic mouse model oral tongues bearing HN31cells following treatment with B02, fractionated radiation dose (5 Gy), and in combination as described in Materials and Methods. J and K, Kaplan-Meier analysis for survival and relative body weight loss during the treatment with B02 and radiation in HN31 tumor bearing mice respectively. Each treatment group contains 5-6 mice. All in vivo data were expressed as ± standard error mean (±SEM) and P values < 0.05 were considered significant.