Figure 3.

piR-4447944 promotes migration and invasion, and inhibits apoptosis of PCa cells. A, Wound-healing assay evaluated the migration ability of LNCaP cells and 22Rv1 cells with piR-4447944 overexpression. B-C, Transwell assays confirmed that the migration (B) and invasion (C) ability of PCa cells was enhanced with piR-4447944 overexpression. D-E, LNCaP cells (D) and 22Rv1 cells (E) transfected with NC or piR-4447944 mimic were cultured in an androgen-deprived medium for 48 h. The medium contained 10% CS-FBS and corresponding IC₅₀ of enzalutamide (around 20 μM enzalutamide for LNCaP, and 50 μM enzalutamide for 22Rv1). Then apoptosis of PCa cells was evaluated by staining with Annexin V/PI, followed by flow cytometry analysis. The histogram showed the percentage (%) of early and late apoptotic cells from three independent experiments. F, Western blot detection of cleaved caspase 3 protein in control and piR-4447944-overexpressed LNCaP cells and 22Rv1 cells with or without ADT (medium containing 10% CS-FBS + the corresponding IC₅₀ of enzalutamide). Results are presented as mean ± SD. *p < 0.05; **p < 0.01, ***p < 0.001, ****p < 0.0001. Data were obtained from at least three independent experiments.