Mechanistic investigation of artesunate against head and neck squamous cell carcinoma through network pharmacology and experimental validation.

BACKGROUND: Artesunate (ART), a semisynthetic derivative of artemisinin, has demonstrated antitumor activity in multiple malignancies; however, its cytotoxic activity and underlying mechanisms in head and neck squamous cell carcinoma (HNSCC) remain incompletely understood. METHODS: Network pharmacology and molecular docking were employed to predict potential molecular targets of ART in HNSCC. The effects of ART on cell viability, migration, invasion, and apoptosis were evaluated in SCC-25 and CAL27 cells in vitro. Expression changes in the Bcl-2/Bax/Caspase-3 axis were assessed using Western blotting and RT-qPCR. A xenograft model was further established to examine the in vivo antitumor activity of ART. RESULTS: Network pharmacology analysis identified 77 potential ART-associated targets related to HNSCC. GO and KEGG enrichment analyses suggested that these targets were involved in several cancer-related pathways. PPI analysis highlighted six core targets, among which CASP3 exhibited the most favorable predicted interaction with ART in docking analysis. Experimental results showed that ART treatment was associated with reduced cell viability, migration, and invasion, along with increased apoptotic activity in HNSCC cells. These effects were accompanied by an increased Bax/Bcl-2 ratio and elevated cleaved Caspase-3 levels. In the xenograft model, ART treatment suppressed tumor growth and increased tumor cell apoptosis. CONCLUSION: ART demonstrates preclinical antitumor activity in HNSCC and is associated with activation of Bcl-2/Bax/Caspase-3 axis. These findings provide preliminary mechanistic insight and support further investigation of ART in preclinical models of HNSCC.