Figure 3: miR-96 and miR-182 reduce migration and invasion of breast cancer cells in vitro.

Migration and invasion abilities were evaluated using wound healing, transwell migration, and Matrigel invasion assays. Hs578 cells were transfected by hsa-miR-96, hsa-miR-182 or pcDNA3 control plasmid (Ctrl). Wound healing assay results (a) showed migration inhibition by hsa-miR-182. Pictures were taken at the indicated time points after scratch. The graph to the right represents the width of remaining open wound calculated in relation to time 0 (n=3). (b) Overexpression of hsa-miR-96 and hsa-miR-182 reduced invasion as was demonstrated by Matrigel invasion assay. Photos of representative fields are on the left. Results were calculated as invasion rate in relation to control cells. Transwell migration assay (c) and Matrigel invasion assay (d) of 4T1 cells stably expressing mmu-miR-96 or mmu-miR-182 showed decreased migration and invasion abilities compared with 4T1 cells stably expressing scrambled sequence as control (Ctrl). Photos of representative fields are on the left. Results were calculated as migration or invasion rates in relation to control cells (n=3). (e) Wound healing assay showing increased migration following reduction of hsa-miR-182 or hsa-miR-96. MCF-7 cells were transfected by antago-miR-182, antago-miR-96 or scrambled control. Pictures were taken at the indicated time points after scratch. The graph to the right represents the width of remaining open wound calculated in relation to time 0 (n=4). Downregulation of mmu-miR-96 and mmu-miR-182 by antago-miR transfection enhanced migration (f) and invasion (g) of 4T1 cells. Scale bars, 50 μm. Values are presented as mean±s.e.m. (Student’s t-test, *P<0.05, **P<0.01, ***P<0.005).