Figure 3.

MiR-199a-5p and miR-214-3p inhibitors could block phenotypic transition and migration but increase adhesion of HG-HPMCs in vitro. (A) Real-time PCR shows the level of miR-199a-5p or miR-214–3p in HG-HPMCs after transfection with the miR-199a-5p or miR-214–3p inhibitor or the control oligonucleotide (inc). Mannitol-treated HPMCs were followed as osmotic pressure control. (B) Expression of E-cadherin, claudin-2, α-SMA, CTGF, and fibronectin in HG-HPMCs transfected with the miR-199a-5p or miR-214–3p inhibitor was evaluated by western blot analysis and compared with the control oligonucleotide. β-actin was used as a loading control. (C) Immunohistochemical analysis of E-cadherin and claudin-2 expression in HG-HPMCs transfected with the miR-199a-5p or miR-214–3p inhibitor compared with the control oligonucleotides. Original magnification, ×400. (D) Real-time PCR showing the mRNA levels of E-cadherin, claudin-2, α-SMA, CTGF, and fibronectin transfected with the miR-199a-5p or miR-214–3p inhibitor compared with the control oligonucleotide. (E) Cell migration assays were performed using HG-HPMCs transfected with the miR-199a-5p inhibitor, miR-214–3p inhibitor, or control oligonucleotides. Mannitol-treated HPMCs were followed as osmotic pressure control. Representative fields of migrated HG-HPMCs on the membrane. Original magnification, ×200. Average numbers of migrated cells in three independent experiments. P values were determined using t tests. (F) Cell adhesion assays. Bar graphs represent the average adhesive numbers of cells on the Matrigel after 0.5, 1, 2, and 4 hours ±SEM. P values were determined using t tests. *P<0.05; **P<0.01.