Dingxiang Guanshitong suppresses ESCC progression by modulating TAK1/NF-κB/GLUT1-associated glycolytic reprogramming

Esophageal squamous cell carcinoma (ESCC) is a highly aggressive tumor with poor clinical outcomes and limited treatment options. Dingxiang Guanshitong (DGST), a traditional Chinese herbal formulation, has shown potential activity in ESCC, but its molecular mechanisms require further clarification. The current study aimed to evaluate the therapeutic impact and underlying biological actions of DGST on ESCC. Network pharmacology was used to identify candidate DGST constituents and targets, followed by KEGG pathway enrichment analysis. Antitumor effects of DGST was examined in a 4-nitroquinoline 1-oxide (4NQO)-induced ESCC mouse model using H&E staining, immunohistochemistry, TUNEL assay, and flow cytometry. Proteomics analysis identified differentially expressed proteins and enriched pathways. In TE-1 cells, TAK1 knockdown/overexpression stable lines were constructed by lentiviral transduction, and pathway involvement was evaluated using immunofluorescence, luciferase reporter assays, CCK-8, Transwell, scratch assays, qRT-PCR, western blotting, and metabolic indicator assays. DGST showed dose-associated suppression of ESCC tumor progression, cell proliferation, migration, and invasion, while promoting apoptosis. Proteomics suggested that DGST affected glycolysis and NF-κB-related pathways. DGST treatment was associated with reduced phosphorylation of TAK1, IκBα, and p65, lower GLUT1, HK2, PFKP, and LDHA expression, decreased ATP levels, and increased MPC1 as well as mitochondrial complex indicators. TAK1 knockdown enhanced the effects of DGST, whereas its overexpression reversed these effects. DGST may inhibit ESCC progression by modulating TAK1/NF-κB/GLUT1 axis-mediated glycolytic reprogramming. The findings elucidate the potential therapeutic value of DGST in the treatment of ESCC and provide mechanistic insights for its clinical application.