Osthole and matrine suppress malignant progression of lead-exposed NSCLC via regulating cellular homeostasis: A combined network pharmacology and in vitro experimental study

Network pharmacology and docking identified key compounds and targets. • Osthole and matrine were key bioactives against lead-exposed NSCLC. • Core targets and pathways in lead-driven NSCLC were identified. • In vitro assays confirmed inhibition of NSCLC progression. Background Non-small cell lung cancer (NSCLC) is the predominant subtype of lung malignancy. Accumulating epidemiological evidence demonstrates that environmental lead (Pb) exposure as a critical driver of its initiation and progression. Purpose This study integrated network pharmacology, molecular docking, and in vitro assays to investigate the oncogenic effects of low-concentration Pb on A549 cells and explore the potential therapeutic mechanisms of osthole and matrine. Study design and methods Network pharmacology identified key potential signaling pathways, and molecular docking screened genes with higher binding affinity to the target. MTT assay was used to determine CC 50 , maximum safe concentration (MNTC) and cellular proliferative capacity of Pb, matrine and osthole. Scratch test and Matrigel invasion test detected A549 cell migration and invasion; flow cytometry detected apoptosis, ∆Ψm, ROS and cell cycle. qRT-PCR and Western blot measured gene expression levels. Results Network pharmacology analysis showed that the shared targets of osthole, Pb and NSCLC were mainly enriched in the PI3K/AKT and apoptosis pathways, while matrine, Pb, and NSCLC targets were primarily enriched in cancer metabolism, key hub genes included CDK2, CASP9, IKBKB, PDPK1 and MDM2. Molecular docking demonstrated that osthole showed strong binding to HSD11B1 and CDK2, while matrine exhibited high affinity for DRD4 and CDK2. Considering the essential functions of CDK2 in G1 phase transition, PDPK1, IKBKB, and CASP9 in cell survival, inflammatory signaling, and mitochondrial apoptosis, and MDM2 in p53-mediated cell cycle regulation, phenotypic validation experiments were conducted. Pb exposure significantly enhanced malignant characteristics of A549 cells, including increased migratory and invasive abilities, decreased apoptosis, and a significant reduction in mitochondrial membrane potential. At the molecular level, Pb exposure disrupted cellular homeostasis, as reflected by altered expression of SIRT-1, NRF-1, TGF-β1, Smad, STAT3, STAT1, and mtTFA. Treatment with osthole or matrine effectively reversed all lead-induced abnormalities. Conclusion Taken together, osthole or matrine exerts anti-cancer effects against Pb- promoted NSCLC progression by restoring disrupted cellular homeostasis through modulating cell cycle progression, inhibiting Pb-induced mitochondrial dysfunction and stress-related signaling imbalance. Download: Download high-res image (130KB) Download: Download full-size image