Figure 3.

Amlexanox represses the metastasis of PCa cells in vivo. (A-C) Pretreatment with Amlexanox significantly suppresses the metastases formation of PC3-m cells (Amlexanox group: n=6; DMSO group: n=5). Quantitative analysis of the total photon flux is shown in Figure 3C (mean of each metastasis). (D) Immunochemical staining of human nuclei in sections of mouse femurs. Human nuclei were stained by the anti-nuclei antibody (Merck MAB1281, clone 235-1) that only recognizes the human-specific nucleus to identify the human prostate cancer cells in the mouse xenograft models. Positively stained cells are indicated by arrowheads. Scale bar = 20 μm. (E) Amlexanox does not exert toxicity when administered systemically to mice even at a high dosage of 30 mg/kg. Different doses of Amlexanox were injected intra-peritoneally to nude mice twice a week for 2 weeks to see toxicity. Alterations in body weight was not detected upon Amlexanox treatment. (F) Amlexanox treatment resulted in a decrease of the in vivo tumor-initiating ability of PC3 cells by the limited dilution assay. Unpaired t-test was used for the statistical analysis. *, P<0.05. Data are presented as mean ± SEM.