Fusobacterium nucleatum FadA promotes the progression of esophageal squamous cell carcinoma through binding to TFRC

In China, esophageal squamous cell carcinoma (ESCC) represents the major histopathological form of esophageal cancer. Fusobacterium nucleatum (F. nucleatum, Fn) has been reported to be associated with ESCC. FadA has been identified exclusively in F. nucleatum. However, the potential role and underlying mechanism of FadA in ESCC remain unclear. Whole genome sequencing (WGS) data of ESCC tissue from NCBI database were analyzed to characterize the microbial composition of ESCC. The relative expression of Fn and FadA was validated by qPCR in another ESCC tissue cohort. An E. coli BL21 (DE3) expression system was utilized to express FadA. After purification, FadA was co-cultured with KYSE150 cells. Cell migration, proliferation, and invasion were evaluated by wounding-healing, CCK-8, and Transwell migration and invasion assays. RNA and proteomic sequencing analysis were performed to identify the altered molecule initiated by FadA treatment. TFRC was further validated by knockout and overexpression in KYSE150 cells. Molecular docking, molecular dynamics simulations, and normal mode analysis (NMA) were used to predict and evaluate the interaction between FadA and TFRC. ESCC tissue exhibited decreased α-diversity and distinct microbial composition, with significant enrichment in F. nucleatum. FadA expression was significantly elevated in ESCC tissue samples. Functional assays illustrated that FadA promoted the migration, proliferation, and invasion of ESCC cells. Integrated RNA and proteomic sequencing identified TFRC as a key upregulated target after FadA treatment, which was further confirmed by TFRC overexpression and knockout. Computational analysis supported a stable interaction between FadA and TFRC. FadA is upregulated in ESCC and promotes the migration, proliferation, and invasion in ESCC through TFRC. FadA may serve as a potential biomarker and therapeutic target in ESCC.