Fig. 1

A flow chart of the experimental design. First, to explore the newly identified miRNAs in HBV-positive HCC patients, five pairs of HCC and adjacent matched non-HCC tissue from HBV-positive and HBV-negative patients were collected using high-throughput sequencing. Bioinformatics analyses were performed to predict the biological function of significant differentially expressed miRNAs. Second, the online TCGA database was mined for miRNAs related to HBV-positive HCC, and a bioinformatics analysis of these miRNAs was also performed. Third, differentially expressed miRNAs with overlapping expressionpatterns in both our miRNA-seq data and the TCGA database were chosen for further bioinformatics analysis. First, to validate the high-throughput sequencing results, a few novel miRNAs and miRNAs with overlapping expressionpatterns were randomly chosen for qRT-PCR analysis in 15 pairs of HCC samples with or without HBV infection. Meanwhile, a survival analysis was conducted using a few miRNAs with overlapping expressionpatterns from the TCGA database. Finally, functional experiments were performed, including CCK-8, cell cycle transition, apoptosis, and luciferase reporter assays to further uncover the miRNA’s underlying mechanisms in HBV-positive HCC