Fig. 6.

Proglumide decreases metastases in the mT5 orthotopic mouse model (n = 8 per group) after 21 days. A: final mT5 tumor weights after 21 days: Mean tumor weights ± SE of orthotopic mT5 tumors show that dietary fat significantly increases the size of tumors in mice on a high-fat diet compared with a regular diet (*P = 0.023). The tumors of mice treated with proglumide (prog) were smaller than the mice on untreated (reg) water but this did not reach significance. B: mean number of metastases per mouse was significantly increased in mice fed a high-fat diet compared with those on a regular diet (**P = 0.007). Proglumide significantly reduced the number of metastases in mice on a high-fat diet (***P = 0.004). C: Masson’s trichrome stain in representative tumor from each treatment group shows increased fibrosis in mice on either high-fat or control (cont) diet but with untreated drinking water (left). Proglumide treatment decreased the fibrosis in both groups of mice receiving either high-fat or control diets (right). D: quantitative analysis of fibrosis scoring shows significantly decreased fibrosis in both groups of mice treated with proglumide (*P < 0.02) compared with mice on regular untreated drinking water. E: immunoreactivity for αSMA is shown (cat. no. Ab-124964, Abcam, Cambridge, MA; 1:3,500) in a representative tumor from a mouse on the high fat diet and regular drinking water (bottom left, n = 10) and a representative image showing αSMA immunoreactivity in tumors of mice on the high-fat diet that received proglumide in the drinking water (bottom right, n = 10). Quantitative analysis of αSMA-positive tissue does not reveal any significant difference in tumors of mice on proglumide (P = 0.288). F: results of metastases PCR array showing genes from tumors with significant fold-changes when mice were placed on a high-fat diet compared with regular diet. αSMA, alpha smooth muscle actin. αSMA, alpha smooth muscle actin; mT5, murine mT5-2D cells; NS, not significant.