Multiple targets and pathways non-monotonically regulate lung squamous cell carcinoma migration in response to...

Phthalates (PAEs) are widely used plasticizers that are increasingly linked to tumorigenesis. In this study, we integrated network toxicology and machine learning to elucidate the potential mechanisms of PAEs in lung squamous cell carcinoma (LUSC). By integrating multi-database target prediction, The Cancer Genome Atlas Program (TCGA) transcriptomic analysis, and algorithmic screening, four key genes, ANXA5, CDK4, MMP1, and SRD5A1, were identified as major targets of PAEs. Molecular docking and dynamics simulations indicated stable binding of butyl benzyl phthalate (BBP) to ANXA5, and SRD5A1. In vitro validation using H226 and NCI-H1703 cells confirmed these predictions. BBP at non-cytotoxic concentrations (0.1-10 μM) altered cell behavior: 0.1 μM BBP promoted migration and upregulated SRD5A1 and ANXA5, whereas 10 μM BBP downregulated ANXA5, showing a non-monotonic "low-dose activation-high-dose inhibition" pattern typical of endocrine disruptors. Collectively, these results indicate that BBP interferes with LUSC progression through multiple target and multipathway mechanisms, including direct binding to key proteins, disturbance of steroid metabolism, and modulation of membrane signaling. This integrative study links environmental phthalate exposure with lung cancer pathogenesis and provides a hypothesis worthy of further investigation for risk assessment and targeted prevention.