https://orcid.org/0000-0002-2482-5619
Abstract
Objective
The mechanisms of pseudouridine synthase (PUS) are not definite in hepatocellular carcinoma (HCC), the objective of this study is to investigate the effect of PUS genes in HCC.
Materials and Methods
Differentially expressed and prognostic gene of PUS enzymes was identified based on The Cancer Genome Atlas (TCGA), International Cancer Genome Consortium (ICGC) and Gene Expression Profiling Interactive Analysis (GEPIA) databases. For the identified gene, pseudouridine synthase 1 (PUS1), was used for further research. The clinicopathological feature of PUS1 was analyzed by Student’s t-test. Prognostic significance was explored by Kaplan-Meier (KM) analysis and Cox proportional hazards regression model. Receiver operating characteristic (ROC) curve was applied to appraise diagnostic and prognostic value. The Database for Annotation, Visualization, and Integrated Discovery (DAVID) and Gene Set Enrichment Analysis (GSEA) were implemented to explore mechanism of PUS1. A Guangxi cohort was applied to verify differential expression. In vitro cell experiments were implemented to investigate the influence for proliferation, reactive oxygen species (ROS) level, migration, and invasion of HCC cells after a knockdown of PUS1.
Results
PUS1 was significantly overexpressed in HCC tissues, and patients with high PUS1 were related to unpromising clinicopathological features. Survival analysis revealed high PUS1 expression was associated with a poor overall survival (OS) and 1 year-recurrence free survival (RFS), was an independent risk factor. Meanwhile, ROC curve showed that PUS1 had a diagnostic and prognostic significance to HCC. Functional enrichment analysis implied that PUS1 may be involved in metabolic pathways, mitochondrial function, non-alcoholic fatty liver disease (NAFLD), and some important carcinogenic pathways. Cell assays revealed that knockdown of PUS1 significantly constrained the migration, proliferation, invasion and improved the ROS level of HCC cells.
Conclusion
PUS1 may be a prognostic biomarker and a underlying treatment target for HCC.
Abbreviations
PUS, pseudouridine synthase; TCGA, The Cancer Genome Atlas; ICGC, International Cancer Genome Consortium; HCC, hepatocellular carcinoma; MST, median survival time; RFS, recurrence-free survival; OS, overall survival; HR, hazard ratio; CI, confidence interval; AFP, alpha fetoprotein; PIVKA-II, Protein Induced by Vitamin K Absence or antagonist-II; GALAD, gender, age, AFP-L3, AFP and des-carboxy-prothrombin; HBV, hepatitis B virus; HCV, hepatitis C virus; GEPIA, Gene Expression Profiling Interactive Analysis; ROC, receiver operating characteristic; DAVID, Database for Annotation, Visualization and Integrated Discovery; GO, gene ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, Gene Set Enrichment Analysis; PCR, polymerase chain reaction; WB, Western blot; ROS, reactive oxygen species; NAFLD, non-alcoholic fatty liver disease.
Data Sharing Statement
The data used to support the findings of this study are included within the article and the Supplementary Information Files.
Ethics Approval and Informed Consent
This study has been authorized by the Ethical Review Committee of the First Affiliated Hospital of Guangxi Medical University [Approval Number: 2023-E071-01]. Written informed consent was agreed and signed by all HCC patients. This study complies with the Declaration of Helsinki.
Acknowledgments
The authors thank the contributors of TCGA, ICGC and GSE14520 database for sharing the HCC dataset on open access. We greatly acknowledge the support of the National Key Clinical Specialty Programs (General Surgery and Oncology) and the Key Laboratory of Early Prevention & Treatment for Regional High-Incidence-Tumor (Guangxi Medical University), Ministry of Education, China. We sincerely appreciate the helpful comments on this study received from editor and reviewers. We are grateful to Zaida Huang, Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region, People’s Republic of China, for his help with data collection, data analysis, technical assistance, formatting-related writing assistance in this paper.
Author Contributions
All authors contributed to data analysis, drafting or revising the article, have agreed on the journal to which the article will be submitted, gave final approval of the version to be published, and agree to be accountable for all aspects of the work.
Disclosure
The authors report no conflicts of interest in this work.