Figure 2

miR-3127-5p affects proliferation, migration and motility of A549 and H292 cells.

(A) and (B): Left panel: Q-PCR determination of miR-3127-5p expression in A549 cells and H292 cells stably transfected with Lv-GFP, Lv-3127, or Lv-3127-off. Right panel: Growth curves of (A) A549 cells and (B) H292 cells stably transduced with control Lv-GFP; Lv-3127, a vector for overexpression of miR-3127-5p; or Lv-3127-off, a vector for expression of an miR-3127-5p inhibitor. Cells (1000/well) were cultured in a 96-well plate and cell growth was monitored every 24 h for 7 days using a CCK-8 assay, each type of cell was analyzed in quadruplicate (p < 0.01 for A549 cells and H292 cells, repeated measure ANOVA). (C): Cell cycle analysis. The percentage of cells in G0/G1 phase increased and that in S phase decreased in Lv-3127 transduced A549 (left panel) and H292 (right panel) cells; the opposite was observed in Lv-3127-off transduced cells. (D): miR-3127-5p overexpression reduced cell motility and miR-3127-5p knockdown increased cell motility in the wound scratch assay in both A549 (left panel) and H292 (right panel) cells. A uniform scratch was made in each confluent layer culture; the extent of the wound closure was monitored under a phase-contrast microscope and photographs were taken at 0, 12 and 24 h. Triplicate experimentation generated similar results. (E): A549 (upper panels) and H292 (lower panels) cells were loaded onto the top well of the transwell inserts in triplicate for cell migration assay. Photographs are representative of cells that had migrated to the bottom chamber after 48 h. Cells were stained with hematoxylin and observed under a microscope (×200). Invasion was quantified by determining the total number of cells that had migrated through the membrane.