Abstract
Neutrophils expelling neutrophil extracellular traps (NETs) during the process of NETosis play a critical role in tumor development. In this study, we aimed to identify a prognostic NETosis-related gene signature that could predict the immunetherapy response and explore the expression of these genes in hepatocellular carcinoma (HCC). Differentially expressed NETosis-related genes were identified from the GSE80489 dataset in the Gene Expression Omnibus (GEO), and a protein-protein interaction (PPI) network was constructed using Cytoscape and the STRING database. Gene Ontology (GO) enrichment analysis revealed associations with chemical stimuli and the positive regulation of cellular processes in HCC. A total of 37 differentially expressed NETosis-related genes were identified, and SNORA31, PMAIP1, GLA, CXCL1, and ADM were selected to construct a prognostic signature. In the training cohort, the low-risk group demonstrated significantly better overall survival (OS) compared to the high-risk group (P = 2.715e-06), and multivariable analysis confirmed that this signature was an independent prognostic factor for HCC (HR = 1.630, 95% CI = 1.295-2.052, P < 0.001). This signature also significantly predicted OS in validation cohorts (P = 9.704e-03) and was associated with HCC patients' OS according to the Kaplan-Meier Plotter. Moreover, it could predict the immunotherapy response in HCC patients. Finally, we investigated the expression of NETosis-related genes in single-cell subtypes and in the HepG2 cell line, providing further insights into their relevance in HCC. Overall, this NETosis-related gene signature serves as a prognostic indicator and a potential predictor for immunotherapy response in HCC.