ZEB1-upregulated LCN2 via transcription regulation affects ferroptosis and malignant progression in non-small cell lung cancer.

Abstract

Ferroptosis exerts a remarkable influence on the progression of non-small cell lung cancer (NSCLC). Although the contribution of lipocalin 2 (LCN2) to NSCLC pathogenesis has been demonstrated, further elucidation of the molecular determinants driving LCN2 dysregulation is essential for developing NSCLC interventions. mRNA levels were performed by quantitative polymerase chain reaction (PCR), and protein expression was assessed by immunoblotting and immunohistochemical assays. Reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and Fe^2 + levels were measured to analyze cell ferroptosis. The relationship between zinc-finger E-box-binding homeobox 1 (ZEB1) and the LCN2 promoter was verified by chromatin immunoprecipitation (ChIP) and luciferase assays. Our data showed that LCN2 and ZEB1 levels were upregulated in NSCLC. LCN2 depletion reduced the growth, motility, and invasiveness of NSCLC cells, while promoting apoptosis and ferroptosis. LCN2 depletion also inhibited the tube formation of human umbilical vein endothelial cells (HUVECs) in vitro. Mechanistically, ZEB1 enhanced the transcription and expression of LCN2 in NSCLC cells. ZEB1 downregulation diminished HUVEC tube formation, suppressed the growth, motility, and invasiveness of NSCLC cells, and enhanced apoptosis and ferroptosis. Notably, these effects were counteracted by re-expression of LCN2. Additionally, ZEB1 downregulation inhibited the growth of xenograft tumors in vivo. Our study demonstrates the pro-tumorigenic role of the ZEB1/LCN2 cascade in NSCLC by promoting malignant progression and impeding ferroptosis. Such molecular insights may help devise novel candidates for NSCLC treatment.