Figure 7.

PI3k/AKT-dependent activation of NF-κB by miR-520c-3p is important for HBx-induced migration and invasion

(A) miR-520c-3p activates NF-κB (p-IκBα). Huh7 cells were pretreated with 5 mM NF-κB inhibitor SC-514 or dimethyl sulfoxide for 4 h followed by transfection with NC or miR-520c-3p mimic. After 48 h, the levels of the indicated proteins were examined by western blot analyses. (B) HBx induces cell migration and invasion via activating NF-κB (p-IκBα). Huh7 cells were pretreated with SC-514 followed by transfection with pxj40-HA-HBx or empty vector. They were then subjected to western blot analyses for the indicated proteins (left) or cell migration and invasion assays as in Figure 2A (right). (C and D) Inhibition of the NF-κB pathway via inactivation of IκBα reverses the ability of miR-520c-3p or HBx to induce EMT and cell invasion. Huh7 were co-transfected with MIGR1-IκBα and miR-520c-3p or HBx for 48 h, and then the levels of the indicated proteins were examined by western blot analysis (C and D, left), and the cell migration and invasion assays were performed (D, right). (E) miR-520c-3p activates NF-κB (p-IκB). Huh7 cells were pretreated with signaling inhibitors (PD98058 for MAPK/ERK, SP600125 for JNK, XAV-939 for Wnt, rapamycin for mTOR, and AS1842856 for FOXO) for 4 h followed by transfection with NC or miR-520c-3p mimic and subjected to invasion assays. Three independent experiments were performed, and representative data are shown. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 in (B–D).