The online version of this article (https://doi.org/10.1186/s13046-018-0681-y) contains supplementary material, which is available to authorized users.
Hepatitis B virus (HBV) plays a critical role in the tumorigenic behavior of human hepatocellular carcinoma (HCC). MicroRNAs (miRNAs) have been reported to participate in HCC development via the regulation of their target genes. However, HBV-modulated miRNAs involved in tumorigenesis remain to be identified. Here, we found that a novel highly expressed miRNA, TLRC-m0008_3p (miR-3928v), may be an important factor that promotes the malignancy of HBV-related HCC.
Solexa sequencing was applied to profile miRNAs, and RT-qPCR was used to identify and quantitate miRNAs. We studied miR-3928v function in HCC cell lines by MTT, colony formation, migration/invasion, and vascular mimicry (VM) assays in vitro and by a xenograft tumor model in vivo. Finally, we predicted and verified the target gene of miR-3928v by a reporter assay, studied the function of this target gene, and cloned the promoter of miR-3928v and the transcription factor for use in dual-luciferase reporter assays and EMSAs.
A variant of miR-3928 (miR-3928v) was identified and found to be highly expressed in HBV (+) HCC tissues. Voltage-dependent anion channel 3 (VDAC3) was validated as a target of miR-3928v and found to mediate the effects of miR-3928v in promoting HCC growth and migration/invasion. Furthermore, HBx protein increased early growth response 1 (EGR1) expression and facilitated its translocation into the nucleus to enhance miR-3928v promoter activity in an NF-κB signaling-dependent manner.
miR-3928v is induced by HBx through the NF-κB/EGR1 signaling pathway and down-regulates the tumor suppressor gene VDAC3 to accelerate the progression of HCC.
Additional file 1: Supplementary methods: Detailed experimental materials and methods. Figure S1. Plasmid transfection efficiency. Figure S2. VDAC3 promotes the apoptosis of HCC cells. Figure S3. HBV/HBx infection enhances the malignancy of HCC cells via miR-3928v. (DOCX 1489 kb)13046_2018_681_MOESM1_ESM.docx
Additional file 2: Table S1. Sequences of primers used in this study. (DOCX 29 kb)13046_2018_681_MOESM2_ESM.docx
Additional file 3: Table S2. Novel miRNAs identified by Solexa sequencing. (DOCX 30 kb)13046_2018_681_MOESM3_ESM.docx
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- miR-3928v is induced by HBx via NF-κB/EGR1 and contributes to hepatocellular carcinoma malignancy by down-regulating VDAC3
- BioMed Central
Journal of Experimental & Clinical Cancer Research
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