This study aims to investigate the ferroptosis-inducing effects of Tianma Granules (TMG) in colorectal cancer and elucidate its molecular mechanisms. Ferroptosis, an iron-dependent form of regulated cell death, represents a novel therapeutic target for cancer. We combined network pharmacology with experimental validation to explore TMG’s anti-cancer potential through ferroptosis modulation.

Network pharmacology identified 382 ferroptosis-related genes overlapping with 12,944 CRC-associated targets (p < 0.05), with SLC7A11, GPX4, SAT1, PTGS2, and GLS2 prioritized as core targets. In vitro, TMG dose-dependently suppressed CRC cell proliferation (p < 0.05), elevated reactive oxygen species (p < 0.05) and ferrous ion levels (p < 0.01), effects reversed by ferroptosis inhibitor Ferrostatin-1. c-Casp3 levels were unchanged (p > 0.05), excluding apoptosis Transmission electron microscopy revealed mitochondrial cristae fragmentation and vacuolation, hallmark features of ferroptosis. Molecular analyses demonstrated TMG-mediated downregulation of SLC7A11 and GPX4, alongside upregulation of SAT1, PTGS2, and GLS2 (p < 0.05). In xenograft models, high-dose TMG (23.2 g/kg) reduced tumor volume, attenuated cachexia, and elevated intratumoral ROS and Fe2+ levels (p < 0.01), corroborating ferroptosis induction in vivo.

TMG suppresses CRC progression by inducing ferroptosis via dual inhibition of SLC7A11/GPX4 and activation of SAT1/PTGS2/GLS2. This study bridges traditional medicine and ferroptosis biology, positioning TMG as a novel therapeutic candidate for CRC.