Hepatitis B virus X protein (HBx)-mediated immune modulation and prognostic model development in hepatocellular carcinoma

PLoS One. 2025 Jun 27;20(6):e0325363. doi: 10.1371/journal.pone.0325363. eCollection 2025.

Abstract

Hepatitis B virus (HBV) X protein (HBx) is critical in hepatocellular carcinoma (HCC) development, but its influence on tumor immunity and the tumor microenvironment (TME) remains unclear. This study aimed to construct a prognostic model based on HBx-related genes and explore their relationship with immune infiltration and immunotherapy response. Through transcriptome sequencing of our HBx-expressing HepG2 cells and analysis of HCC patient data from the cancer genome atlas (TCGA) and genotype-tissue expression (GTEx), we identified seven HBx-related genes, nuclear VCP-like (NVL), WD repeat domain 75 (WDR75), NOP58 nucleolar protein (NOP58), Brix domain-containing protein 1 (BRIX1), deoxynucleotidyltransferase terminal interacting protein 2 (DNTTIP2), MKI67 FHA domain interacting nucleolar phosphoprotein (NIFK), and ribosome production factor 2 (RPF2), associated with poor prognosis. LASSO Cox regression narrowed these to four key genes (BRIX1, RPF2, DNTTIP2, and WDR75), which were used to develop a prognostic riskscore signature. High-risk patients exhibited lower survival rates, decreased infiltration of anti-tumor immune cells, poorer responses to immunotherapy, and increased immune evasion. Among the four genes, DNTTIP2 showed higher expression in single-cell data, was linked to migration inhibitory factor (MIF) signaling, and may play a pivotal role in shaping an immunosuppressive TME. Elevated DNTTIP2 expression was confirmed in HBx-expressing HepG2 cells and HBV-infected HCC samples. This study highlights a novel HBx-related four-gene prognostic model that predicts clinical outcomes, immune infiltration, and immunotherapy response, offering insights into HCC progression and potential therapeutic targets.

MeSH terms

  • Carcinoma, Hepatocellular* / genetics
  • Carcinoma, Hepatocellular* / immunology
  • Carcinoma, Hepatocellular* / mortality
  • Carcinoma, Hepatocellular* / pathology
  • Carcinoma, Hepatocellular* / virology
  • Female
  • Gene Expression Regulation, Neoplastic
  • Hep G2 Cells
  • Hepatitis B virus
  • Humans
  • Liver Neoplasms* / genetics
  • Liver Neoplasms* / immunology
  • Liver Neoplasms* / mortality
  • Liver Neoplasms* / pathology
  • Liver Neoplasms* / virology
  • Male
  • Prognosis
  • Trans-Activators* / genetics
  • Trans-Activators* / immunology
  • Trans-Activators* / metabolism
  • Tumor Microenvironment / immunology
  • Viral Regulatory and Accessory Proteins

Substances

  • Viral Regulatory and Accessory Proteins
  • hepatitis B virus X protein
  • Trans-Activators