Panoply™ Human CCL20 Over-expressing Stable Cell Line

Given the relatively low uptake of human papillomavirus (HPV) vaccines and the limited treatment options for existing cervical cancer cases, there is an urgent need to develop more precise prognostic models. Furthermore, the clinical significance of circadian clock characteristics in cervical cancer remains under-explored. In this study, researchers successfully stratified cervical cancer patients into high-risk and low-risk groups based on circadian clock oscillation patterns. Subsequently, they constructed a prognostic risk model comprising GJB2, CCL20, and KRT24, which demonstrated excellent predictive value for overall survival (OS). Analyses revealed that metabolic imbalances-particularly within the glycolysis pathway-and immune-related pathways were the primary distinguishing features between the high- and low-risk groups. Further investigation showed that the tumor microenvironment (TME) of the high-risk group exhibited an "immune-desert" phenotype characterized by the infiltration of suppressive myeloid cells, a pattern associated with resistance to immunotherapy. Finally, the study confirmed a positive correlation between CCL20 levels and clinical stage in patient cohorts; in mouse models, CCL20 expression led to a significant reduction in CD8+ T-cell infiltration and a marked increase in M2-like macrophage infiltration.

Here, the researchers established a stable CCL20-overexpressing model (OE-CCL20) in U14 cells and confirmed successful overexpression via qRT-PCR, Western blot, flow cytometry, and immunohistochemistry (IHC). Subsequently, they established a U14 subcutaneous tumor model in C57BL/6 mice. Although no accelerated tumor growth was observed in the CCL20-OE group (Figure 1H), the mice in this group exhibited significantly poorer survival outcomes. Flow cytometric analysis of the tumor microenvironment (TME) revealed a higher M2/M1 macrophage ratio in the CCL20-overexpression group (Figure 1E, F). Concurrently, this group showed reduced numbers of CD8+ T cells, decreased CD107a expression and increased PD-1 expression on CD8+ T cells, while the proportion of Treg cells remained unchanged; these findings were consistent with the CIBERSORT analysis results (Figure 1F, G). Finally, given the significantly poorer prognosis observed in high-risk patients within the immunotherapy cohort, the researchers treated CCL20-OE mice with anti-PD-L1 therapy and similarly observed resistance to immunotherapy (Figure 1H). This further confirms the presence of an "immune-desert" tumor microenvironment in this mouse model, characterized by a reduction in CD8+ T cells and an increase in M2-type macrophages.

Figure 1. CCL20 as an independent indicator for worse prognosis and capable of enhancing M2 macrophages infiltration. (Yu Y, et al., 2023)