Figure 2. Effect of SAM on MDA-MB-231 tumor growth and metastasis.

(A) Schematic representation of SAM-treatment in MDA-MB-231 tumor xenograft mice. Female CD1 mice inoculated with MDA-MD-231-GFP cells via orthotopic route were randomized, and treatment with SAM at different doses was started from day three post tumor cell inoculation. Animals were treated daily with vehicle alone or SAM (40 mg/kg/day or 80 mg/kg/day) via daily oral gavage. (B) Tabular representation of the incidence of tumor in control and two experimental groups. (C) Tumor volume was determined at weekly intervals from week 5 when the animals started to develop tumors. Treatment with SAM caused a significant dose-dependent decrease in tumor growth. Results are representative of mean ± SEM of tumor volumes obtained from at least seven animals per group. Significant differences were determined using ANOVA followed by post hoc Bonferroni test and are represented by asterisks. (^**P < 0.01; ^***P < 0.001). (D) To evaluate the effect of SAM on tumor metastasis, control and SAM (40 and 80 mg/kg/day) treated animals were sacrificed at week 10 and different organs (lung, liver, spleen) were collected. Organ slices of 1-mm thickness were mounted on a glass slide, and the GFP-positive foci were examined under the fluorescent microscope. Ten randomly selected slides were counted and averaged to determine the GFP-positive metastatic foci in each organ. Significant differences were determined using ANOVA followed by post hoc Bonferroni test and are represented by asterisks. (^*P < 0.05; ^**P < 0.01, and ^***P < 0.001).