Effects of BBP on cell migration and invasion
(A) Huh7 cells were transfected with two different AhR (top) and NF-κB (bottom) shRNA as described in the Methods, and then protein levels were detected by immunoblotting. (B) Transwell migration and invasion assays. Huh7 cells transfected with control, AhR, or NF-κB shRNAs were seeded onto inserts, treated with or without BBP, and allowed to migrate for 24 (migration) or 48 (invasion) hours. The numbers of migrating and invading cells were counted under a microscope. Scale bars: 100 μm. (C) Tumor growth of intrahepatically injected Huh7-IFP cells in vehicle control and BBP treatment groups for the indicated times of was detected by non-invasive imaging. (D) Numbers of other organs with metastasis of vehicle control or BBP treatment group (n = 18 for each group) after 4 weeks. (E) Immunohistochemical staining for PI3K and NF-κB in the tumor region of the mouse liver after treatment of BBP for 4 weeks. Magnificantion, 40x. The intensity of PI3K and NF-κB staining was analyzed by Tissue Quest software. Data shown in panels B and E are representative of three independent experiments. Each value is the mean ± SD of three independent experiments. The asterisks indicate a significant difference between vehicle treated group and BBP treated groups, as analyzed by Student’s t-test (*p < 0.05; **p < 0.01).