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. 2014 Nov 6;6(1):258–270. doi: 10.18632/oncotarget.2724

Figure 3. Resistin promotes cell migration and matrix metalloproteinase (MMP-2) expression by down-regulating microRNA (miR)-519d expression.

Figure 3

(A) The JJ012 and SW1353 cells were transfected with miR-519d mimic or inhibitor for 24 h, and cell migration ability was examined by Transwell assay. (B) The JJ012 cells were transfected with an miR-519d mimic or inhibitor for 24 h, and MMP-2 expression was examined by western blot (upper panel), and enzyme-linked immunosorbent assay (lower panel). (C) The JJ012 and SW1353 cells were incubated with resistin (0.3–3 ng/ml) for 24 h, and miR-519d expression was detected by real-time quantitative polymerase chain reaction. (D) Sequences of miR-519d and the potential miR-519d binding site at the MMP-2 3′ untranslated region (3′ UTR; WT-MMP-2 3′ UTR). Also shown are the nucleotides mutated in the MMP-2 3′UTR mutant (MUT-MMP2 3′ UTR). (E) The JJ012 and SW1353 cells were transfected with a wild-type or mutant MMP-2 3′ UTR luciferase plasmid for 24 h followed by stimulation with resistin (0.3-3 ng/ml) for 24 h, and the relative luciferase activity was measured. The JJ012 cells were pre-treated with AMPK and p38 inhibitors for 30 min or pre-transfected with specific siRNAs for 24 h followed by stimulation with resistin (3 ng/ml) for 24 h; and the (F, G) miR-519d expression and (H) MMP-2 3′ UTR activity were examined. The results are expressed as mean ± SEM. *, P < 0.05 compared with control. #, P < 0.05 compared with the resistin-treated control group.