Adamantyl Retinoid-Related Molecules Induce Apoptosis in Pancreatic Cancer Cells by Inhibiting IGF-1R and Wnt/β-Catenin Pathways
Figure 4
Dose-response effect of 3-Cl-AHPC on CD44+/CD24+ cells sphere formation and apoptosis in PANC-1 sphere cells. (a) Addition of 0.25, 0.5, and 1.0 μM 3-Cl-AHPC added at time of cell seeding inhibited sphere formation at 7 and 14 days. (b) 0.5 and 1.0 μM 3-Cl-AHPC inhibited sphere formation when added 7 days following sphere formation. (c) 1.0 μM 3-Cl-AHPC induced apoptosis in CD44+/CD24+ sphere cells as indicated by nuclear fragmentation detected by acridine orange/ethidium bromide and (d) DAPI staining. Spheres were visualized and photographed utilizing a fluorescence microscope. (e) Apoptosis of sphere cells as demonstrated by TUNEL assay. CD44+/CD24+ spheres were treated with 1.0 μM ARRs for 7 days (7D) after sphere formation. Details of slides preparation, visualization, antibodies utilized, and TUNNEL assay methodologies were as described in Section 2.