Exploration of the Values of Macrophage Differentiation-Related Genes inPrognosis and Immunotherapy Prediction for Hepatocellular Carcinoma

INTRODUCTION: Hepatocellular carcinoma (HCC) poses significant global health and economic challenges, with tumor-associated macrophages (TAMs) implicated in its progression. This study aimed to develop a prognostic signature based on macrophage differentiation-related genes (MDRGs) for HCC risk stratification. MATERIALS AND METHODS: Transcriptomic data from HCC and adjacent tissues were analyzed to identify differentially expressed genes (DEGs). Prognostic MDRGs were screened via univariate Cox regression and refined using LASSO regression. A multivariate Cox-derived risk model was established and validated using Kaplan-Meier survival analysis. Immune infiltration and immunotherapy responses were assessed, alongside in vitro functional validation of key genes. RESULTS: Analysis identified 67 prognostically relevant MDRGs from HCC-specific DEGs. LASSO regression and multivariate Cox analysis yielded a robust 9-MDRG signature. Patients stratified into high-risk groups exhibited significantly shorter survival, elevated macrophage infiltration, and enhanced predicted responses to anti-PD1/CTLA4 therapies. In vitro studies revealed that NCAPG, a transcription factor within the signature, promotes HCC progression by upregulating the core regulator EZH2. DISCUSSION: The established 9-MDRG signature effectively stratifies HCC patients into distinct risk groups with significant prognostic divergence. Its strong association with heightened TAM infiltration underscores the biological relevance of macrophage differentiation in HCC progression. Crucially, the model exhibits predictive value for markers associated with immunotherapy response, suggesting high-risk patients may benefit preferentially from antiPD1/CTLA4 regimens. Furthermore, the mechanistic dissection of the NCAPG-EZH2 axis within the signature unveils a key molecular pathway linking macrophage biology to HCC aggressiveness, nominating NCAPG and EZH2 as novel therapeutic targets. CONCLUSION: We developed a clinically translatable 9-MDRG prognostic model that effectively predicts HCC outcomes and informs immunotherapy strategies. The identified NCAPG-EZH2 axis highlights potential therapeutic targets for macrophage-mediated HCC progression.