Effects of PRL-3-WT/mutations and prenylation inhibitor FTI-277 on B16F1 melanoma cells adhesion and migration ability.

<p>(A) B16F1 cells transfected with wild type or mutated Myc-PRL-3 expression vectors and (B) PRL-3-WT transfected cells pretreated with 1 µM or 2 µM FTI-277 for 18 hours were seeded into the wells of 96-well plates coated with fibronectin (10 µg/ml). After incubation for 30 minutes at 37°C, non-adherent cells were gently washed away. The amount of the adherent cells was determined at 592 nm after crystal violet staining. Data are mean±SEM of three independent experiments and each experiment includes triplicate sets. (C) A single-cell suspension (100 µl, 1×10⁶ cells/ml) of cells was plated into the upper wells of Transwell inserts containing 8 µm pore polycarbonate membranes pre-coated with fibronectin on the under surface. Cells were allowed to migrate for 24 hours at 37°C, and then fixed, stained with crystal violet and counted. Quantitative analysis of the number of the cells migrated to the lower side of the membrane is shown. (D) Treatment with FTI-277 inhibits the migration of B16F1 melanoma cells. B16F1 cells were transfected with PRL-3-WT and incubated with FTI-277 1 µM or 2 µM for 18 hours before plated into the upper wells of Transwell inserts, which still contain different concentrations of FTI-277. All Data are mean±SEM of three independent experiments. *, P<0.05; **, P<0.01, vs. mock vector for (A) and (C) and vs. PRL-3-WT expression vector for (B) and (D).