Figure 3.

miR-218-5p Promotes Trophoblast Invasion, Migration, and EVT Outgrowth

(A–F) Cells were seeded on transwells without and with Matrigel to assess cell migration or invasion, respectively. Stable transfection of mir-218-1 increased cell migration (A) and invasion (B) compared with control cells transfected with EV. Transient transfection of the miR-218-5p mimics also promoted cell migration (C) and invasion (D) compared with NC. Conversely, anti-miR-218-5p significantly decreased the migratory (E) and invasive (F) ability of the cells compared with controls. The results presented in (A)–(F) are data pooled from a minimum of three independent experiments. (G) Explants from first-trimester placentas were placed on Matrigel and treated with miR-218-5p or control (200 nM) for 24 hr. EVT outgrowth was measured at the time of treatment (0 hr) and at termination of the experiment (24 hr). An increase in EVT outgrowth was observed in miR-218-5p-treated tissues. (H) Explant tissues were treated with anti-miR-218-5p or the control (200 nM) for 48 hr. A decrease in EVT outgrowth was observed with anti-miR-218-5p treatment. Neither miR-218-5p (G) nor anti-miR-218-5p (H) had significant effects on cytotoxicity. Representative images are shown, and bar graphs present data from four experiments with unique placentas. *p < 0.05, **p < 0.01, ***p < 0.001. Error bars represent SEM. Scale bars, 250 μm.