Transcriptomic identification of HBx-associated hub genes in hepatocellular carcinoma

Zhengzhong Ni; Jun Lu; Weiyi Huang; Hanif Khan; Xuejun Wu; Danmei Huang; Ganggang Shi; Yongdong Niu; Haihua Huang

doi:10.7717/peerj.12697

Transcriptomic identification of HBx-associated hub genes in hepatocellular carcinoma

PeerJ. 2021 Dec 22:9:e12697. doi: 10.7717/peerj.12697. eCollection 2021.

Authors

Zhengzhong Ni^#¹, Jun Lu^#², Weiyi Huang¹, Hanif Khan¹, Xuejun Wu¹, Danmei Huang¹, Ganggang Shi¹, Yongdong Niu¹, Haihua Huang³

Affiliations

¹ Department of Pharmacology, Shantou University Medical College, Shantou, Guangdong, China.
² Department of Hepatobiliary Surgery, First Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong, China.
³ Department of Pathology, Second Affiliated Hospital of Shantou University Medical College, Shantou, Guangdong, China.

^# Contributed equally.

Abstract

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies around the world. Among the risk factors involved in liver carcinogenesis, hepatitis B virus (HBV) X protein (HBx) is considered to be a key regulator in hepatocarcinogenesis. Whether HBx promotes or protects against HCC remains controversial, therefore exploring new HBx-associated genes is still important.

Methods: HBx was overexpressed in HepG2, HepG2.2.15 and SMMC-7721 cell lines, primary mouse hepatocytes and livers of C57BL/6N mice. High-throughput RNA sequencing profiling of HepG2 cells with HBx overexpression and related differentially-expressed genes (DEGs), pathway enrichment analysis, protein-protein interaction networks (PPIs), overlapping analysis were conducted. In addition, Gene Expression Omnibus (GEO) and proteomic datasets of HBV-positive HCC datasets were used to verify the expression and prognosis of selected DEGs. Finally, we also evaluated the known oncogenic role of HBx by oncogenic array analysis.

Results: A total of 523 DEGs were obtained from HBx-overexpressing HepG2 cells. Twelve DEGs were identified and validated in cells transiently transfected with HBx and three datasets of HBV-positive HCC transcription profiles. In addition, using the Kaplan-Meier plotter database, the expression levels of the twelve different genes were further analyzed to predict patient outcomes.

Conclusion: Among the 12 identified HBx-associated hub genes, HBV-positive HCC patients expressing ARG1 and TAT showed a good overall survival (OS) and relapse-free survival (RFS). Thus, ARG1 and TAT expression could be potential prognostic markers.

Keywords: HBx; Hepatocellular carcinoma; Hub genes; Transcriptomic.

Grants and funding

This study was funded by the National Natural Science Foundation of China (81772972, 81572703), Natural Science Foundation of Guangdong Province (2021A1515010776), Science and Technology Planning Project of Guangdong Province Special Project Fund (210728166902052), Department of Education, Guangdong Government under the Top-tier University Development Scheme for Research and Control of Infectious Diseases (2016026, 2015060, 2015089). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.