ZG16 Overexpression Curbs Colorectal Cancer Cell Growth and Boosts Macrophage Phagocytosis in Lab Tests

Scientists probing the molecular machinery of colorectal cancer have turned their attention to ZG16, a gene whose activity appears to fade as the disease takes hold. In a study published in BMC Medical Genomics, Fan Sun, Kai-Li Liao, Xia-Jing Yu and Xiao-Zhong Wang combined large-scale genomic data analysis with laboratory experiments on SW620 colorectal cancer cells to examine what happens when ZG16 is forced back into action. Their findings, drawn from bulk RNA sequencing, single-cell sequencing and cell-based assays, suggest that restoring ZG16 expression may blunt several malignant behaviors of cancer cells while simultaneously making them more visible to the immune system’s scavenger cells.

The research team began by mining bulk RNA sequencing data from The Cancer Genome Atlas to assess ZG16 expression across colorectal cancer samples. Gene Set Enrichment Analysis, Gene Set Variation Analysis and immune infiltration analysis were applied to this dataset. After correcting for cell composition using deconvolution algorithms, the investigators analyzed ZG16 expression in relation to patient survival. The results showed that ZG16 was downregulated in colorectal cancer, yet the gene lacked independent prognostic value, meaning its expression level alone did not predict patient outcomes once other factors were accounted for.

To gain finer resolution, the team integrated single-cell data from the Gene Expression Omnibus repository. This analysis revealed that ZG16 is mainly expressed in goblet and epithelial cells, and that its activity is relatively lower in malignant epithelial cells than in non-malignant subsets. Within ZG16-positive epithelial cells, the researchers observed an inverse correlation between ZG16 and two molecules: CD47 and CD24. These molecules are known to act as signals that help cancer cells evade phagocytosis, the process by which macrophages engulf and destroy abnormal cells. The inverse relationship between ZG16 and these anti-phagocytic markers was associated with enhanced phagocytic recognition and hemophagocytosis scores, alongside suppressed negative regulatory pathways.

With these computational signals in hand, the researchers moved to the laboratory bench. They overexpressed ZG16 in SW620 colorectal cancer cells and assessed the consequences using a battery of functional assays. Cell Counting Kit-8 assays, flow cytometry and transwell chambers were employed to measure proliferation, apoptosis, migration and invasion. The results were consistent: ZG16 overexpression inhibited proliferation, migration and invasion, while inducing apoptosis, the programmed cell death pathway that cancer cells typically suppress.

The team then examined whether the gene’s influence extended to immune recognition. They overexpressed ZG16 in SW620 cells and evaluated macrophage phagocytosis via flow cytometry, while fluorescence microscopy visualized the close physical association between macrophages and the cancer cells. ZG16 overexpression reduced the expression of CD47 and CD24 on the cancer cells and, critically, increased macrophage-mediated phagocytosis in vitro. This dual effect, lowering the molecular shields that cancer cells use to avoid immune clearance while simultaneously boosting the actual engulfment by macrophages, points to a mechanism by which ZG16 might influence the tumor microenvironment beyond its direct effects on cell behavior.

The authors conclude that ZG16 was downregulated in colorectal cancer and predominantly expressed in goblet and 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. These observations position ZG16 as a gene whose loss may contribute to both the aggressive growth of colorectal cancer cells and their ability to dodge immune surveillance.

Several caveats frame the significance of this work. The functional experiments were conducted entirely in vitro, using a single colorectal cancer cell line, SW620. Whether ZG16 overexpression would produce similar effects in other colorectal cancer cell lines, in animal models or in patients remains untested in this study. The bulk RNA sequencing analysis also found that ZG16 lacked independent prognostic value, so the gene’s expression level did not serve as a standalone predictor of survival in the patient cohort examined. The single-cell findings, while revealing a clear inverse correlation between ZG16 and CD47/CD24 within ZG16-positive epithelial cells, describe associations rather than demonstrated causal relationships at the single-cell level.

The study’s methodological approach reflects a growing trend in cancer genomics research: layering bulk transcriptomic data with single-cell resolution to identify genes whose activity patterns suggest functional importance, then validating those signals through direct perturbation experiments. By correcting for cell composition in the bulk data using deconvolution algorithms, the researchers aimed to distinguish true gene expression changes from shifts in the proportions of different cell types within tumor samples. The integration of ligand-receptor interaction analysis from single-cell data further allowed the team to connect ZG16 expression patterns to known immune signaling pathways.

Colorectal cancer remains a major global health challenge, and the molecular regulators driving its progression are still poorly defined despite extensive research. Genes like ZG16, which is predominantly expressed in goblet cells, a specialized epithelial cell type that secretes mucus in the intestinal lining, may play roles that conventional bulk tissue analysis obscures. The present study’s finding that ZG16 expression inversely correlates with CD47 and CD24, two well-characterized anti-phagocytic signals, offers a plausible molecular link between the loss of this goblet cell gene and the ability of malignant epithelial cells to resist immune clearance.

The work is an early-access publication, shared ahead of the final Version of Record, and is subject to further edits. It is citable and carries a permanent DOI. As with all cell-line-based studies, the findings represent a starting point for further investigation rather than a clinical recommendation. Future research would need to determine whether ZG16’s effects on malignant behavior and macrophage phagocytosis hold across diverse colorectal cancer models and whether manipulating ZG16 expression could offer a therapeutic avenue. For now, the study adds to the evidence that ZG16, a gene tied to goblet cell identity, may act as a brake on colorectal cancer cell aggressiveness and as a facilitator of immune-mediated clearance in laboratory settings.

Subject of Research: The role of ZG16 overexpression in colorectal cancer cell behavior and macrophage phagocytosis in vitro

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

Article References: Sun, F., Liao, K.-L., Yu, X.-J., & Wang, X.-Z. (2026). Potential role of ZG16 overexpression in malignant behaviors and macrophage phagocytosis in vitro. BMC Medical Genomics. https://doi.org/10.1186/s12920-026-02478-4

Image Credits: AI Generated

DOI: 10.1186/s12920-026-02478-4

Keywords: ZG16, colorectal cancer, SW620 cells, macrophage phagocytosis, CD47, CD24, apoptosis, goblet cells, single-cell sequencing, The Cancer Genome Atlas, tumor immunology, in vitro