Lactylation of β-catenin promotes non-small cell lung cancer by inhibiting ferroptosis through TCF4-mediated GPX4 transcription

Ferroptosis is an iron-dependent form of regulated cell death driven by the accumulation of lipid peroxides. Lactylation, a post-translational modification derived from metabolic reprogramming, whether it drives non-small cell lung cancer (NSCLC) progression by regulating ferroptosis remains unclear. This study aims to elucidate the role and underlying mechanism by which β-catenin lactylation regulates ferroptosis to drive NSCLC progression, offering novel insights into disease pathogenesis and potential therapeutic targeting. Blood, tumor tissues, and adjacent normal tissues from NSCLC patients were collected to detect lactate levels and protein lactylation using biochemical assays, Western blot, and immunofluorescence. Cell models were established in BEAS-2B and NSCLC cell lines, with lactylation regulated by lactate (LA) and 2-deoxyglucose (2-DG). Cellular proliferation, migration, and invasion were quantified using CCK-8, EdU incorporation, Transwell chamber, and wound healing assays, respectively. Ferroptosis was assessed by detecting LPO and GSH levels, and transmission electron microscopy. β-catenin lactylation sites were identified by mass spectrometry and site-directed mutagenesis. β-catenin, TCF4, and GPX4 expression at protein and mRNA levels was detected by Western blot and qRT-PCR. ChIP verified the binding of β-catenin/TCF4 complex to the GPX4 promoter. Nude mouse subcutaneous xenograft models were used to validate in vivo tumor growth regulation, with immunofluorescence detecting β-catenin lactylation and GPX4 expression in tumors. NSCLC clinical samples showed elevated lactate levels, global protein lactylation, and β-catenin lactylation at lysine 193 (K193) compared to controls. β-catenin K193 lactylation enhanced protein stability, promoting NSCLC cell proliferation, migration, and invasion, which was reversed by 2-DG. LA accelerated tumor growth in nude mice, while 2-DG inhibited it. Mechanistically, β-catenin K193 lactylation reduced LPO and increased GSH to suppress ferroptosis: lactylated β-catenin translocated to the nucleus, formed a complex with TCF4, and upregulated GPX4 transcription by binding its promoter. GPX4 overexpression reversed ferroptosis induced by β-catenin K193 lactylation inhibition, while blocking β-catenin/TCF4 interaction downregulated GPX4 and enhanced ferroptosis. A systematic literature search was conducted across international English-language databases including PubMed, Web of Science, Scopus, and Medline, covering publications from database inception through October 2024. The core search terms, including “non-small cell lung cancer (NSCLC)”, “β-catenin”, “lactylation”, “ferroptosis”, “GPX4”, and “TCF4”, were combined to identify studies exploring the interplay among metabolic modifications, signaling pathways, and regulated cell death mechanisms in NSCLC. Following initial retrieval, non-research articles such as reviews, conference abstracts, and case reports were excluded, along with duplicate publications. Priority was given to original experimental studies published within the past decade that included in vitro (cell-based) or in vivo (animal model) investigations, as well as analyses of clinical NSCLC specimens. Emphasis was placed on studies providing mechanistic insights into the “metabolic modification–signaling pathway–cell death” axis, particularly those offering quantifiable molecular data or validated experimental outcomes. A final set of 111 core references was selected to substantiate the mechanistic hypotheses and analytical framework of this study.