Figure 4.

MTDH is a direct target gene of miR-375. (A) The 3′-UTR of MTDH mRNA includes a highly conserved binding site for miR-375. (B) HEK293 cells were co-transfected with pYr-MTDH-3U and an miR-375 mimic or NC. At 24 h after transfection, luciferase activity was detected. (C) Dual-luciferase assays demonstrated miR-375 mimic-mediated repression of pYr-MTDH-3U luciferase; however, no significant difference from pYr-MTDH-3Umt luciferase was identified. (D) miR-375 modulated the expression of MTDH, cyclin D1, VEGF-C and E-cadherin in EC109 cells. β-Actin served as a loading control. Experiments were performed in triplicate and data are expressed as the mean ± standard deviation, **P<0.01 vs. NC. MTDH, metadherin; miR-375, microRNA-375; 3′-UTR, 3′-untranslated region; NC, negative control; mt, mutated; VEGF-C, vascular endothelial growth factor.