Identification of ADH4 from a ferroptosis-oxidative stress prognostic model in hepatocellular carcinoma

Background Ferroptosis is a recently identified form of regulated cell death that is closely associated with oxidative stress and redox homeostasis. Although ferroptosis-related prognostic models have been developed for hepatocellular carcinoma (HCC), the prognostic value of integrating oxidative stress-related features with ferroptosis-associated alterations remains unclear. Methods We identified oxidative stress- and ferroptosis-related genes (OFRGs) and developed a prognostic signature for HCC using transcriptomic and clinical data from The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) cohorts. Consensus clustering based on OFRG expression profiles revealed distinct molecular subgroups with different survival outcomes. Kaplan-Meier and ROC analyses assessed the prognostic performance. GO, KEGG, Tumor Mutation Burden and Immune Checkpoints and Tumor Microenvironment analyses uncovered underlying molecular pathways. Functional experiments including CCK-8, Clone Formation Tests, Wound Healing Assay and Western Blotting further validated these mechanisms. Results A three-gene prognostic model consisting of AKR1B15, S100A9, and ADH4 was established using Cox regression and LASSO analyses. The risk score effectively stratified HCC patients according to overall survival in internal validation cohorts, while external validation demonstrated moderate predictive performance, particularly for short- and intermediate-term survival prediction. The risk score was also associated with differences in immune infiltration patterns, immune checkpoint expression, and predicted drug sensitivity. Among the signature genes, ADH4, previously reported to be associated with HCC prognosis, was downregulated in HCC tissues and independently correlated with patient survival. Functional assays showed that ADH4 overexpression inhibited HCC cell proliferation and migration in vitro. Conclusions Collectively, this study provides an oxidative stress- and ferroptosis-associated prognostic framework for HCC and identifies ADH4 as a candidate biomarker with preliminary functional evidence, requiring further mechanistic validation.