Integrated bioinformatics to identify and validate the role of oxidative stress-related gene EPHX2 in laryngeal cancer.

Laryngeal cancer is a highly aggressive malignancy with high incidence and mortality rates. This study aims to identify potential biomarkers and therapeutic targets for laryngeal cancer through an integrated approach combining bioinformatics and machine learning, followed by experimental validation. We obtained laryngeal cancer tissue sample data from The Cancer Genome Atlas (TCGA) database and used Weighted Gene Co-expression Network Analysis (WGCNA) to identify key gene modules. These modules were then intersected with oxidative stress-related genes from the GeneCards database to determine core genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of the core genes revealed that they are primarily involved in responses to xenobiotic stimuli, oxidative stress, and multiple signaling pathways. Cox regression analysis identified GPT and EPHX2 as independent risk factors for laryngeal cancer. Through a comprehensive analysis using LASSO regression and Protein-Protein Interaction (PPI) networks, highlighted EPHX2 as a candidate gene of interest. Experimental results showed that the expression of EPHX2 in laryngeal cancer tissues was significantly lower than that in control tissues (p < 0.01). Additionally, EPHX2 expression was associated with increased infiltration of M0 macrophages (p < 0.01) and decreased infiltration of CD4 memory T cells (p < 0.01), suggest that EPHX2 expression correlates with immune-cell infiltration patterns and clinical outcomes. Western blot and immunohistochemistry corroborated the differential expression of EPHX2 between tumor and non-tumor tissues. Collectively, these findings uncover an oxidative stress-related gene signature in laryngeal cancer and suggest that EPHX2 expression correlates with immune-cell infiltration patterns and clinical outcomes. However, the present data are observational and do not establish a mechanistic or causal role for EPHX2 in laryngeal cancer progression or immune modulation; functional studies such as gene knockdown or overexpression assays are required to validate its biological role.