FIGURE 7.

Caffeic acid suppresses the clonogenicity and pluripotency of colorectal CSCs by disrupting Akt signaling in a mice xenograft model. Schematic representation of the experimental protocol as described in the section “Materials and Methods.” Anesthetized 7 weeks old male NSG mice were inoculated with a 1:1 mixture of Matrigel and 3 × 10⁶ HCT116 cells into the subcutaneous tissue. Mice bearing HCT116 cell tumors were treated with caffeic acid (10 mg/kg, intraperitoneally) or vehicle (PBS). The tumor volumes (A) and weights (B) were measured as described in the section “Materials and Methods.” Non-treated and caffeic acid-treated colorectal cancer tissues were stained with antibodies specific for CD44 and CD133. DAPI staining was used to label the nuclei within each field (C). The relative activation levels of Akt signaling in the HT29 cell xenograft models were assessed by western blotting (D). Non-treated and caffeic acid-treated colorectal cancer tissues were stained with an antibody specific for phospho-Akt. DAPI staining was used to label the nuclei within each field (E). β-actin was used as the internal control. The data represent the mean ± SD of three independent experiments.