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. 2020 Jan 15;9:1535. doi: 10.3389/fonc.2019.01535

Figure 2.

Figure 2

miR-20b-5p suppresses migration and invasion of PC3 and DU145 cells by regulating EMT-related gene expression. (A) qRT-PCR detected the expression of miR-20b-5p in PC3 and DU145 cells transfected with a miR-20b-5p mimic or control mimic for 24 h. ***P < 0.001 vs. control mimic. Migration (B) and invasion (C) assays for PC3 and DU145 cells transfected with a miR-20b-5p mimic and treated with 2 ng/mL TGF-β1. Representative images were taken at ×200 magnification; the numbers of migrated cells on the lower surface of the inserts were quantified in four random fields from each treatment group. *P < 0.05, ***P < 0.001 vs. blank control; &P < 0.001 vs. control mimic + TGF-β1; bar = 200 μm. (D) PC3 and DU145 cells were transfected with a miR-20b-5p or control mimic and then treated with or without TGF-β1. Western blot analysis detected E-cadherin, ZO-1, vimentin, and N-cadherin. (E) PC3 and DU145 cells were transfected with miR-20b-5p or control inhibitor and then treated with or without TGF-β1; E-cadherin, ZO-1, vimentin, and N-cadherin expression was detected by western blotting. All experiments were performed in triplicate. (F) Immunofluorescence staining detected the expression of E-cadherin (green) and vimentin (red) in miR-20b-5p mimic-transfected PC3 and DU145 cells. Nuclei were counterstained with DAPI (blue). Scale bars = 25 μm. (G) Phase-contrast microscope observed cell morphology of PC3 and DU145 with miR-20b-5p mimic transfection. Scale bars = 25 μm.