Inhibition of NRF2 driven antioxidant defence and EMT underlies brusatol induced cytotoxicity in cervical cancer cells

Cervical cancer remains a major global health burden, and its therapeutic resistance is strongly influenced by redox adaptation, epithelial-mesenchymal transition (EMT), and metabolic reprogramming. Brusatol, a natural quassinoid compound, is known for its ability to suppress NRF2-driven antioxidant defense mechanisms. However, its mechanistic impact on cervical cancer has not been fully characterized.In this study, we first employed quantitative proteomic profiling to obtain an unbiased overview of Brusatol-induced molecular alterations. Proteomics analysis revealed extensive downregulation of proteins associated with antioxidant defense, glycolysis, cytoskeletal organization, and EMT regulation. Conversely, proteins linked to oxidative stress responses, mitochondrial dysfunction, and apoptotic signalling were significantly upregulated. Pathway enrichment analysis confirmed inhibition of glycolytic and redox-regulatory pathways, suppression of cell motility networks, and activation of oxidative stress driven cytotoxic processes.These global proteomic signatures were further substantiated through western blot validation, which demonstrated reduced expression of key components involved in antioxidant defense and EMT regulation following Brusatol treatment. Western blot analysis also showed suppression of glycolytic markers, supporting the proteomics-predicted metabolic disruption.Functionally, Brusatol significantly reduced proliferation, induced apoptosis, suppressed migration, and reversed EMT characteristics in cervical cancer cell lines. Treated cells exhibited decreased mesenchymal marker expression, restoration of epithelial traits, reduced glucose uptake, and impaired glycolytic activity. Importantly, a non-tumorigenic epithelial cell line showed substantially lower sensitivity, indicating selective cytotoxicity toward cancer cells. Together, these findings demonstrate that Brusatol exerts potent anti-tumor effects in cervical cancer by initiating widespread proteomic reprogramming, suppressing NRF2-linked antioxidant pathways, reversing EMT, and disrupting glycolytic metabolism. The integration of proteomic profiling with biochemical validation and functional assays provides strong mechanistic evidence supporting brusatol as a promising therapeutic candidate for overcoming treatment resistance in cervical cancer.