Homoharringtonine suppresses cervical cancer progression through modulation of the EGFR/PI3K/AKT/mTOR signaling pathway

Abstract

Homoharringtonine (HHT), a natural alkaloid approved for treating hematological malignancies, has shown potential activity against solid tumors, but its role in cervical cancer remains unexplored. This study investigates the anti-cervical cancer effects of HHT and its impact on the EGFR/PI3K/AKT/mTOR signaling pathway. The anti-proliferative, anti-migratory, cell cycle, and pro-apoptotic effects of HHT were evaluated in HeLa and SiHa cervical cancer cells using CCK-8, colony formation, wound healing, Transwell migration, and flow cytometry assays. Protein expression levels of EGFR/PI3K/AKT/mTOR pathway components and their phosphorylated forms were assessed by Western blotting, while network pharmacology, molecular docking, and RNA sequencing were employed to identify potential targets. An in vivo xenograft mouse model was used to evaluate tumor growth inhibition and systemic toxicity. HHT dose-dependently suppressed proliferation, colony formation, and migration of HeLa and SiHa cells, with calculated IC₅₀ values of 62.96 nM and 56.09 nM, respectively, while inducing S-phase arrest and apoptosis. Network pharmacology and transcriptomic analysis pointed to the EGFR/PI3K/AKT/mTOR axis as a key pathway involved, and molecular docking predicted stable binding of HHT to EGFR (-8.8 kcal/mol). Western blotting confirmed that HHT dose-dependently reduced phosphorylation of EGFR (Y1068), PI3K (Tyr458), AKT (Ser473), and mTOR (Ser2448). In the HeLa xenograft model, HHT (0.5 mg/kg/day) significantly inhibited tumor growth without causing body weight loss or major organ toxicity. Collectively, HHT exerts potent anti-cervical cancer activity, with the EGFR/PI3K/AKT/mTOR signaling pathway pointed to as a key mediator of its effects, leading to cell cycle arrest and apoptosis. These findings establish HHT as a promising therapeutic candidate for cervical cancer with a favorable safety profile.