Fig. 4

miR-143, miR-145, and miR-140 inhibit OECM-1 cell proliferation and induce a G2/M arrest. (a) The expression of miR-143, miR-145, and miR-140 is downregulated in the OECM-1 cell line. After transfecting OECM-1 cells with a combination of miR-143, miR-145, and miR-140 at a total concentration of 100nM for 48 h, a significant increase in the expression of these miRNAs can be seen. U6 small RNA was used as internal control, and the relative miRNA levels were analysed using the ddCt method. Error bars represent mean ± SD of three independent experiments. (b) MTT cell proliferation assay was performed after transfecting OECM-1 cells with the 3-miRNA signature. The graph shows the relative cell proliferation rates of untreated controls (C), SC, and transfected (143/145/140) samples. (c) Phases of the cell cycle were analysed based on the PI-stained DNA content of OECM-1 cells transfected with miR-143, miR-145, and miR-140 mimics using a flow cytometric analysis. NC miRNA mimic was used as control. The X-axis of the histogram indicates the intensity of the PI signal and the y-axis represents the cell count. (d) Representative bar graph shows percentage distribution of cells across cell cycle phases. A G2/M phase arrest can be seen in OECM-1 cells transfected with a combination of miR-143/miR-145/miR-140 mimics. Error bars represent mean ± SD of three independent experiments (*p < 0.05, **p < 0.01)