Knockdown of NUAK2 promotes ferroptosis by regulating the mTOR/c-Myc signaling pathway to inhibit endometrial cancer cell progression

Background: Endometrial cancer (EC) represents a major gynecologic malignancy with unknown confirmed causes. Although NUAK family kinase 2 (NUAK2) is overexpressed and has been shown to promote progression in multiple cancers, its role in regulating ferroptosis in EC remains poorly understood. Methods: NUAK2 expression was analyzed using the Gene Expression Profiling Interactive Analysis (GEPIA) database. Western blot detection of protein levels of NUAK2, key ferroptosis regulators (solute carrier family 7 member 11 (SLC7A11) and Glutathione Peroxidase 4 (GPX4)), and major components of the mechanistic target of rapamycin (mTOR)/MYC Proto-Oncogene (c-Myc) signaling pathway. Intracellular ferrous ion (Fe2+) and Reactive Oxygen Species (ROS) levels were quantified to evaluate ferroptosis, small interfering RNA (siRNA)-mediated silencing of NUAK2 and pharmacological interventions, including Ferrostatin-1 (Fer-1) and the mTOR activator MHY1485, were performed to determine the underlying mechanisms. Results: NUAK2 was overexpressed in EC tissues and cell lines. Knockdown of NUAK2 significantly inhibited EC cell proliferation and induced ferroptosis, as indicated by elevated Fe2+ and ROS levels, together with decreased expression of the ferroptosis-related proteins SLC7A11 and GPX4. Mechanistically, NUAK2 knockdown inhibited activation of the mTOR/c-Myc signaling cascade, whereas treatment with the mTOR activator MHY1485 effectively counteracted the ferroptosis induction and proliferation inhibition resulting from NUAK2 silencing. Conclusions: Our findings demonstrate that NUAK2 knockdown promotes ferroptosis and suppresses the malignant progression of EC cells by inhibiting the mTOR/c-Myc signaling pathway. These findings support NUAK2 as a promising therapeutic target in EC treatment.