GSDMD Promotes Epithelial-Mesenchymal Transition and Lymph Node Metastasis in Cervical Cancer in vitro and in vivo
TL;DR
GSDMD dysregulation correlates with activated EMT and lymphangiogenic signaling, potentially facilitating tumor lymphatic dissemination, and is highlighted as a promising therapeutic candidate for restraining cervical cancer metastatic progression.
Abstract
Background Lymph node metastasis (LNM) is a critical determinant of prognosis in cervical cancer (CC). Tumor-associated macrophages (TAMs) and inflammatory signaling within the tumor microenvironment have been implicated in metastatic progression; however, the molecular mechanisms linking inflammation to lymphatic dissemination remain poorly understood. Methods We integrated single-cell RNA sequencing (scRNA-seq) of paired primary tumors and metastatic lymph nodes tissues from a single representative patient with clinical histological validation. Functional roles of gasdermin D (GSDMD) were evaluated using in vitro assays, including proliferation, migration, invasion, ELISA, and RNA sequencing, as well as in vivo xenograft and lymph node metastasis models. Results Single-cell transcriptomic profiling revealed significant activation of pyroptosis-related pathways in metastatic lymph nodes, accompanied by enhanced tumor–stromal interactions. Clinical tissue validation confirmed that GSDMD protein was significantly enriched in metastatic lesions compared to primary tumors. In vitro functional experiments demonstrated that GSDMD promotes epithelial–mesenchymal transition (EMT), enhances migration and invasion, and upregulates VEGFA and VEGFC expression. Mechanistically, GSDMD facilitated the release of pro-inflammatory cytokines such as IL-1β, contributing to a pro-metastatic microenvironment. Notably, GSDMD knockdown suppressed lymph node metastasis in vivo despite promoting primary tumor growth, indicating a context-dependent role in tumor progression. Conclusion Our findings reveal that GSDMD contributes to the pro-metastatic phenotype of cervical cancer. GSDMD dysregulation correlates with activated EMT and lymphangiogenic signaling, potentially facilitating tumor lymphatic dissemination. This study highlights GSDMD as a promising therapeutic candidate for restraining cervical cancer metastatic progression.