Bufothionine inhibits colorectal cancer progression via the STAT3/p53/SLC7A11 signaling axis

Abstract

Background Bufothionine, a major bioactive component of dried toad skin, has shown anti-tumor potential. However, its efficacy and underlying mechanisms in colorectal cancer (CRC) remain unclear. Objective This study aimed to investigate the therapeutic potential of bufothionine and its underlying mechanisms against CRC. Methods Anti-tumor efficacy was assessed using in vitro (CCK-8, migration assays) and in vivo (HT-29 xenograft) models. Network pharmacology, molecular docking and molecular dynamics simulations were employed to predict targets. Mechanisms were validated by biochemical assays, Western blot, IHC, and qPCR, with ferrostatin-1 applied to confirm ferroptosis specificity. Results Bufothionine significantly inhibited HT-29 cell proliferation and migration. Integrated network pharmacology and computational modeling identified p53 and STAT3 as core targets, with potential interactions predicted between bufothionine and key ferroptosis regulators (including SLC7A11). In vitro, bufothionine induced ferroptosis, characterized by increased Fe2+, ROS, and MDA levels, and a decreased GSH/GSSG ratio; these effects were partially reversed by ferrostatin-1. In vivo, bufothionine reduced tumor weight indices, improved liver function by attenuating AST and ALP elevations without hepatosplenic toxicity, and regulated inflammatory cytokines. Mechanistically, bufothionine suppressed inflammatory activation (NLRP3, IL-1β), inhibited STAT3 phosphorylation, and modulated p53 activity, leading to the subsequent inhibition of the SLC7A11/GPX4 axis in both cells and tumor tissues. Conclusion Bufothionine suppresses CRC by the STAT3/p53/SLC7A11 signaling axis in a manner associated with ferroptosis, suggesting its potential as a therapeutic agent for colorectal cancer.