Artemether induces JNK1/FOXO1/GPX4-mediated ferroptosis to suppress colorectal cancer.

BACKGROUND

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, necessitating novel therapeutic strategies. Ferroptosis, an iron‑dependent, lipid-peroxidation-driven form of cell death, has emerged as a potential tumor‑suppressive mechanism. Artemether, a derivative of Artemisia annua L., exhibits antitumor activity in various cancers, but its role in CRC and its link to ferroptosis remain unclear.

METHODS

Using an HCT116 xenograft model and CRC cell cultures, we evaluated artemether's effects on viability, migration, invasion, and histopathology. Inflammatory cytokines (TNF-α, IL-6, IL-1β, IL-18, IL-1, caspase-1) were measured. Ferroptosis was assessed by flow cytometry, electron microscopy, and biochemical markers (GSH, MDA, ROS). Mechanistic studies included network pharmacology, molecular docking, SP600125 treatment, JNK1 siRNA knockdown, ferrostatin-1 rescue, western blotting, and qRT-PCR.

RESULTS

Network pharmacology identified JNK1 as a core target of artemether in CRC. In vivo, artemether dose-dependently suppressed tumor growth, with efficacy comparable to that of oxaliplatin. In vitro, artemether inhibited CRC cell proliferation, migration, and invasion. Concurrently, artemether reduced multiple pro-inflammatory cytokines in serum and cell supernatants, indicating suppression of systemic and local inflammation. Mechanistically, artemether activated the JNK1/FOXO1/GPX4 pathway, leading to Fe²⁺ accumulation, a ROS burst, and lipid peroxidation, thereby inducing ferroptosis. These effects were reversed by the JNK inhibitor SP600125 and JNK1 knockdown. Furthermore, the ferroptosis inhibitor ferrostatin-1 significantly rescued artemether-induced cell death and lipid peroxidation, confirming ferroptosis as the primary mode of cell death.

CONCLUSION

Artemether inhibits CRC and alleviates tumor-associated inflammation by inducing JNK1/FOXO1/GPX4-mediated ferroptosis, validated pharmacologically and genetically, highlighting its potential as an immunomodulatory anticancer agent.