Potential role of ZG16 overexpression in malignant behaviors and macrophage phagocytosis in vitro

Despite extensive research, the key molecular regulators driving Colorectal Cancer (CRC) progression remain poorly defined. We analyzed bulk RNA sequencing (RNA-seq) data from The Cancer Genome Atlas (TCGA) database to assess ZG16 expression, Gene Set Enrichment Analysis (GSEA), Gene Set Variation Analysis (GSVA) and immune infiltration analysis. After correcting for cell composition using deconvolution algorithms, we analyzed ZG16 expression in relation to survival. We integrated single-cell data from Gene Expression Omnibus (GEO) to analyze ZG16 expression, GSVA and ligand-receptor interactions. Furthermore, we assessed the impact of ZG16 overexpression on the proliferation, apoptosis, migration, and invasion of SW620 CRC cells. Finally, we overexpressed ZG16 in SW620 cells and evaluated macrophage phagocytosis via flow cytometry, while fluorescence microscopy visualized their close association. In bulk RNA-seq analysis, ZG16 was downregulated in CRC but lacked independent prognostic value. Single-cell sequencing analysis revealed that ZG16, mainly expressed in goblet/epithelial cells, was relatively lower in malignant epithelial cells than in non-malignant subsets. In ZG16-positive epithelial cells, ZG16 expression inversely correlated with CD47 and CD24, associating with enhanced phagocytic recognition and hemophagocytosis scores but suppressed negative regulatory pathways. Further assessment via CCK-8, flow cytometry, and transwell assays confirmed that ZG16 overexpression inhibited proliferation, migration, and invasion, while inducing apoptosis. Additionally, ZG16 overexpression reduced expression of CD47 and CD24, along with increased macrophage-mediated phagocytosis in vitro. ZG16 was downregulated in CRC and predominantly expressed in goblet/epithelial cells. In SW620 cells, ZG16 overexpression significantly modulated malignant behaviors (suppressing proliferation and invasion, while promoting apoptosis) and increased macrophage-mediated phagocytosis in vitro.