Fig. 4

Mebendazole reduces tumor growth and lung and liver metastases. A Fluorescently labeled MDA-MB-231 cells were implanted into the mammary fat pad of NSG mice. Mice were treated beginning on day 5 with sesame oil as a vehicle gavage and a high-fat KetoCal diet (Con; N = 12), 215 ppm MBZ incorporated into a high-fat KetoCal diet (Feed; N = 7), 20 mg/kg MBZ in a sesame oil gavage (20 mg/kg; N = 6), 30 mg/kg MBZ in a sesame oil gavage (30 mg/kg; N = 12). Mice were exposed to the feed seven days a week, and treatments administered via oral gavage were given four times per week. Tumor volumes are plotted over time. P values for two-tailed t test *< 0.05. B Tumor weights for each treatment group were taken at the endpoint of the experiment (41 days after implantation). P values for two-tailed t test *< 0.05. C Metastatic burden was determined by measuring human genomic HK2 DNA content in the lung of each mouse normalized to their final tumor weight. The final values were normalized by the average of the Con group (mean ± SEM N = 6–12 mice with n = 3 technical replicates). P values for two-tailed t test ****< 0.0001. D Tumor growth curves displaying tumor volume over time prior to tumor removal at Day 39 (mean ± SEM) for NSG mice that began treatment on day 5 with either a high-fat KetoCal diet (Con; N = 11) or 215 ppm of MBZ incorporated into a high-fat KetoCal diet (feed; N = 10). P values for two-tailed t test *< 0.05. E Final tumor weights at the time of tumor removal, 39 days after implantation (mean ± SEM N = 10–11 mice). P values for two-tailed t test *< 0.05. F, G Metastatic burden was determined by measuring human genomic HK2 DNA content in mouse lung F or liver G two-week post-tumor removal surgery. Values were normalized to individual tumor weight and then to the average value of the control group (mean ± SEM, N = 10–11 mice with n = 3 technical replicates). P values for paired two-tailed t test *< 0.05, ****< 0.0001. H Quantification of tumor growth rate (fold change of final tumor volume versus tumor volume at the start of treatment) over two weeks for NSG mice bearing primary HCI-001 PDX tumors. Mice were treated 4 times per week with 30 mg/kg MBZ in sesame oil (N = 9) or sesame oil alone (N = 8) by oral gavage. Two-way paired Student’s t test. I Final tumor weight of HCI-001 PDX primary tumors described in (H). J Metastatic burden was determined as described in (F-G). (Mean ± SEM N = 8–9 mice with n = 3 technical replicates). P values for two-tailed Student’s t test * < 0.05. K Representative lung sections stained with hematoxylin and eosin (H&E) from NSG mice described in 4H. L Quantification of lung metastatic nodules pictured in (K). Mean ± SEM; control (N = 8 mice) and 30 mg/kg MBZ (N = 9 mice). P values for paired two-way t test **< 0.01