Correction to: miR-3928v is induced by HBx via NF-κB/EGR1 and contributes to hepatocellular carcinoma malignancy by down-regulating VDAC3

Abstract

An amendment to this paper has been published and can be accessed via the original article.

Fig. 4

VDAC3 acts as a tumor suppressor. Huh7 and HepG2 cells were transfected with pshR-VDAC3 or controls. HepG2.2.15 cells were transfected with pVDAC3 or controls. Twenty-four hours after transfection, cells were seeded into cell culture inserts. a MTT assays were performed to evaluate cell viability. b Colony formation assays were used to estimate cell proliferation. c Migration/invasion assays and f Western blot analyses were used to evaluate EMT. d VM and g molecular markers of VM were examined to assess tumor angiogenesis. e FACS assays were used to examine changes in cell cycle distribution. (*: P < 0.05)

Fig. 5

VDAC3 is a functional target of miR-3928v. HepG2 cells were co-transfected with pCD3/Flag or pVDAC3 and pri-miR-3928v. RT-qPCR and Western blot analyses (a) and MTT (b), colony formation (c), migration (d), invasion (e) and tube formation assays (f) were performed to analyze whether VDAC3 could functionally rescue the miR-3928v-induced phenotype. (*: P < 0.05, **: P < 0.01, ***: P < 0.001)

Fig. 6

HBx up-regulates miR-3928v expression via EGR1. a Schematic diagram of the location of miR-3928v in the human genome; the relative positions of the miR-3928v promoter constructs are shown (left). Luciferase reporter assays were used to examine miR-3928v promoter activity in Huh7 cells (right). b Promoter activity in HBV (+) and HBV (−) cells (left). Then, the promoter activity in Huh7 cells was examined after HBV or HBx overexpression (right). c The effect of EGR1 on promoter activity (left). The EGR1 binding site in the miR-3928v promoter was mutated, and the luciferase activity induced by EGR1 was examined (right). d ChIP analysis (left) and EMSAs (right) revealed that EGR1 directly binds to the miR-3928v promoter. e The miR-3928v expression level was measured by RT-qPCR after EGR1 overexpression or knockdown. f Blocking EGR1 inhibited HBx-induced miR-3928v promoter activity. g Representative IHC images showing the magnitude of EGR1 expression in tissues isolated from liver cancer patients. h Total and nuclear expression of EGR1 and P65 in Huh7 cells after transfection with HBx was analyzed by Western blot. i An immunofluorescence assay was used to detect the induction of EGR1 expression in the nucleus by HBx in Huh7 cells. j Mechanism by which HBx-induced miR-3928v expression contributes to HCC malignancy. (*: P < 0.05, **: P < 0.01)