Integration Single‐Cell and Spatial Transcriptomics Reveals PRDX1+ Macrophages Colocalize in the Glycolytic Niche and Facilitate Hepatocellular Carcinoma Proliferation and Metastasis Through the PI3K/AKT Signaling Pathway

This study integrated single‐cell RNA sequencing (scRNA‐seq), spatial transcriptomics (ST), and bulk transcriptomics data from public databases to create a comprehensive map of the immune microenvironment in hepatocellular carcinoma (HCC). Cell communication analysis revealed the pivotal role of macrophages in the HCC tumor microenvironment (TME). Through the high‐dimensional weighted gene coexpression network analysis, we identified macrophage‐related gene modules and integrated them with macrophage marker genes from single‐cell sequencing data to obtain intersecting genes. Based on univariate Cox regression analysis and machine learning algorithms, a prognostic model comprising 10 genes was constructed. Through multidimensional analysis, we identified PRDX1 as a core gene associated with macrophage glycolysis pathways. ST data demonstrated that PRDX1 had much higher glycolytic activity. Enrichment analysis revealed that PRDX1‐positive macrophages upregulated pathways related to epithelial–mesenchymal transition (EMT), apoptosis, and hypoxia. Further analysis revealed a complex signaling network among fibroblasts, PRDX1, and T cells. The SCENIC algorithm showed that PRDX1 activated FOXO family transcription factors by increasing the activity of the PI3K/AKT signaling pathway. In vitro experiments confirmed that knocking down PRDX1 significantly decelerated the growth, migration, and invasion of HCC cells while halting the cell cycle and inducing apoptosis. Western blot analysis also showed that PRDX1 may enhance the invasion of HCC cells by accelerating glycolysis and activating the PI3K/AKT signaling pathway. This study elucidates how PRDX1 functions in the HCC TME and provides new theoretical insights and potential therapeutic targets for prognostic assessment and treatment strategies for HCC.