Mechanistic Insights into Dihydroartemisinin's Therapeutic Potential Against Drug-Resistant NSCLC via AURKA Targeting: A Network Pharmacology and Experimental Study

Non-small cell lung cancer (NSCLC) is a malignancy with among the highest incidence and mortality rates globally, and chemotherapy resistance severely limits its therapeutic efficacy. Dihydroartemisinin (DHA), a derivative of artemisinin, exhibits significant potential in antitumor applications, but its molecular mechanism in treating drug-resistant NSCLC remains unclear. This study aimed to investigate the mechanism of DHA in treating cisplatin-resistant NSCLC through the regulation of Aurora kinase A (AURKA). Using network pharmacology, 93 common targets of DHA and NSCLC were identified. Molecular docking analysis revealed strong binding affinity between DHA and AURKA (binding energy: -8.1 kcal/mol), establishing AURKA as a core target. Clinical samples and cellular experiments confirmed that AURKA was significantly overexpressed in lung cancer tissues and A549/DDP-resistant cells, suggesting its close association with cisplatin resistance. In vitro experiments demonstrated that DHA inhibited A549/DDP cell proliferation in a concentration-dependent manner, with IC₅₀ values of 53.14 µM, 36.28 µM, and 23.18 µM at 24, 48, and 72 h, respectively. DHA treatment markedly promoted apoptosis, upregulated the pro-apoptotic protein BAX, downregulated the anti-apoptotic protein BCL-2, and suppressed cell migration and invasion. More importantly, DHA downregulated AURKA expression, activated the AURKA/GSK3β/NRF2 signaling pathway, promoted reactive oxygen species (ROS) accumulation and ferrous iron (Fe²⁺) deposition, and downregulated the expression of key ferroptosis regulators GPX4 and SLC7A11, thereby inducing ferroptosis. Combination experiments revealed a significant synergistic effect between DHA and the AURKA inhibitor VX-680 (combination index CI < 1), with co-treatment further enhancing cytotoxicity against A549/DDP cells and more effectively inducing apoptosis and ferroptosis.A subcutaneous xenograft model in nude mice validated the in vitro findings, showing that DHA combined with VX-680 synergistically inhibited tumor growth. Immunohistochemical staining confirmed significant downregulation of AURKA and GPX4 protein expression in tumor tissues. Safety evaluations indicated no obvious toxic damage to major organs or abnormalities in liver and kidney function indicators in mice treated with either monotherapy or combination therapy.This study is the first to reveal that DHA exerts anti-cisplatin-resistant NSCLC effects by targeting AURKA, activating the AURKA/GSK3β/NRF2 signaling pathway, and simultaneously inducing dual mechanisms of apoptosis and ferroptosis. The combination of DHA with an AURKA inhibitor demonstrates promising synergistic effects and safety, providing a novel potential strategy for clinical treatment of NSCLC resistance and important theoretical support for the development of natural product-based anti-tumor drugs.