<p>Lycorine has been shown to have anti-tumor activity in many cancers, including non-small cell lung cancer (NSCLC). However, its underlying molecular mechanisms still need to be further revealed. The expression levels of spindle component 25 (SPC25) and zinc finger protein 143 (ZNF143) were examined by qRT-PCR and western blot. Cell proliferation, migration and invasion were determined by colony formation assay and transwell assay. Glucose uptake, lactate production and ATP level were evaluated to assess cell glycolysis. The levels of ROS, MDA, GSH and Fe²⁺ were examined to evaluate ferroptosis. Animal experiments were performed to explore the role of SPC25 and lycorine in vivo. The interaction between ZNF143 and SPC25 promoter was confirmed by ChIP and dual-luciferase reporter assays. SPC25 was overexpressed in NSCLC tissues and cells. Lycorine could reduce SPC25 expression. SPC25 overexpression promoted NSCLC cell proliferation, metastasis, glycolysis, suppressed ferroptosis, and enhanced tumorigenesis, while lycorine reversed these effects. ZNF143 could bind to SPC25 promoter region to increase its expression. SPC25 knockdown reversed the promoting effect of ZNF143 on NSCLC cell proliferation, metastasis and glycolysis, as well as the inhibitory effect on ferroptosis, and these effects could be aggravated by lycorine. In conclusion, lycorine may suppress ZNF143/SPC25 axis to inhibit NSCLC malignant progression, providing new evidence that lycorine can be used as an adjuvant therapy for NSCLC.</p> Graphical abstract <p></p>