Anticancer Activity of Fisetin via KEAP1, DNMT1, and mTOR Axis Against Colon Cancer: An Integrated In Silico, In Vitro, and In Vivo Approach

Abstract

The proliferation of colon cancer is influenced by alterations in epigenetic, reactive oxygen species (ROS), and dysregulation of signaling pathways, especially via the involvement of DNMT1, mTOR, and KEAP1 axis. The current investigation examined the antiproliferative capabilities of fisetin (FisT) in a rat model and in colon cancer cell lines HCT116, CT-26, and Caco-2. Our findings show that FisT modulated the DNMT1, mTOR, KEAP1, and IL-6 signaling pathways to enhance the antiproliferative effects against in vitro and in vivo preclinical model. In-silico based anticancer potential of fisetin were revealed through SwissTargetPrediction, Cytoscape, STRING, Schrodinger, molecular dynamics simulations, KEGG, and network analysis. In-vitro MTT assay, cell cycle analysis, ROS level, immunocytofluorescence, western blot analysis, and in vivo immunohistofluorescence, western blot analysis, ELISA-based assays were performed to evaluate anticancer property of fisetin. Our findings revealed that FisT has a strong anticancer property by demethylating DNA, enhancing ROS, KEAP1 level. Simultaneously downregulated the DNMT1 & mTOR expression, and preventing angiogenesis, cell proliferation, invasion, and migration against in vitro & in vivo models. FisT enhances the anticancer response of tumor development and reduces the size of crypt foci. FisT has potential as a cytotoxic drug, improving and intensifying anticancer responses in colon cancer. © 2025 Wiley Periodicals LLC.