Angiotensin—(1–7) and miRNA-149-3p modulates NCI-H460 cell motility and migration.
A) NCI-H460 cells were grown in 6-well plates under different culture conditions. Under confluency, cellular migration was verified by wound healing assay and the recovery area of the injury was measured in each condition analyzed and plotted in graphs. B) Transwell chamber invasion assays of NCI-H460 cells either treated or untreated with the heptapeptide, or transfected with miRNA-149-3p mimics or inhibitor, or in cells cultured in the presence of the ang-(1–7) with subsequent miRNA-149-3p knocked down. Representative field of invaded and stained cells is shown (left), and the measurements were plotted in graphs. In the assays represented in A and B, ANOVA testing showed significant differences between the ability of NCI-H460 cells to recover the injured area of the culture or migrates through the chambers in ang-(1–7) or transfected with miRNA-149-3p mimics groups, compared to the untreated culture (control). C) Relative miRNA expression in logarithmic scale by qRT-PCR. All graphs presents values of the average from three independent experiments, and the error bars represent the standard deviation of the mean. The significance level was set at p<0.05 (***).