Figure 6. The miR186-Twist1 axis suppresses PCa progression.

A. Immunoblotting of epithelial and mesenchymal markers in PC3-Vector, PC3-miR186 and PC3-anti-miR186 cells. B–C, E–F. RTCA monitoring of cell migration (B, E) or invasion (C, F). One group (PC3-Vector, PC3-miR186, PC3-miR186-CD513B and PC3-miR186-Twist1) and another group (PC3-Vector, PC3-anti-miR186, PC3-anti-miR186-pLko.1 and PC3-anti-miR186-shTwist1) cells were seeded into a CIM-Plate without or with pre-coated matrigel (1:40) and subjected to a dynamic analysis lasting for 48 or 96 h, respectively. The migration or invasion slope was shown as histogram. Error bars indicate ±SD, P-values of < 0.001 (***). (also see Supplementary Figure S4). D, G. Soft agar colony formation assays for above two group cells. The culture medium containing 5% FBS with 0.35% agar was layered onto the base. The number of colonies was counted for each well of six-well plates and shown. The experiments were repeated by three independent times with triplicates each. Error bars indicate ±SEM, P-values of < 0.001 (***). H. 1.0×10⁶ of stable cell lines PC3^Luc-Vector, PC3^Luc-miR186, PC3^Luc-miR186-Vector and PC3^Luc-miR186-Twist1 were respectively injected into the left cardiac ventricle of BALB/c nude mice at 7 weeks old by intracardiac injection. Representative bioluminescence images of the micrometastasis was detected by IVIS spectrum image system at 4 weeks after injection and presented visually with various color spots based on different intensity of luminescence.