Background: Aurora kinase A (AURKA) has been reported to play an oncogenic role in non-small cell lung cancer (NSCLC). However, its specific role in regulating mitochondrial function and reactive oxygen species (ROS) in NSCLC cells remains unclear. This study aimed to investigate the function of AURKA in NSCLC, with a particular focus on its regulation of mitochondrial activity and ROS levels in A549 and H1299 cell lines.

Methods: AURKA expression and protein localization in lung adenocarcinoma (LUAD) were analyzed using publicly available gene expression and immunohistochemistry datasets. Small interfering RNA (siRNA) was used to knock down AURKA expression in A549 and H1299 cell lines. To evaluate cellular proliferation, migration, and invasion capabilities, a series of assays were performed, including cell counting kit-8 (CCK-8), colony formation, 5-ethynyl-2′-deoxyuridine (EdU) incorporation, Transwell migration/invasion, and wound healing assays. Mitochondrial function was assessed by measuring mitochondrial membrane potential, nicotinamide adenine dinucleotide/reduced nicotinamide adenine dinucleotide (NAD⁺/NADH) ratio, adenosine triphosphate (ATP) levels, and ROS generation. Apoptosis was evaluated using terminal deoxynucleotidyl transferase dUTP nick end labeling staining, western blot analysis, and caspase activity assays. N-acetylcysteine (NAC) was used in rescue experiments to further investigate the role of oxidative stress.

Results: AURKA was significantly upregulated in LUAD tissues with higher protein expression confirmed by immunohistochemistry data, correlating with poor prognosis (p < 0.05). Targeted inhibition of AURKA expression resulted in diminished proliferative, migratory, and invasive behaviors in NSCLC cells (p < 0.05). AURKA silencing induced mitochondrial dysfunction, decreased NAD⁺/NADH ratio and ATP production, and increased ROS levels, resulting in enhanced apoptosis (p < 0.05). NAC treatment partially reversed these effects (p < 0.05).

Conclusion: AURKA maintains mitochondrial integrity and redox homeostasis in NSCLC cells, thereby suppressing ROS-mediated apoptosis. Targeting AURKA and oxidative stress pathways may represent a promising therapeutic strategy for NSCLC.