Acetyl tributyl citrate promotes osteosarcoma progression via EGFR/AKT signaling: an integrated network toxicology and experimental study

Introduction Acetyl tributyl citrate (ATBC), a widely used plasticizer, has raised health concerns, yet its role in osteosarcoma (OS) remains unclear. This study aimed to investigate whether ATBC promotes OS progression and to explore the underlying molecular mechanisms. Methods An integrated approach combining network toxicology, molecular dynamics simulations, and functional experiments was used. Database mining and protein–protein interaction network analysis were performed to identify and prioritize candidate targets. Functional assays and epidermal growth factor receptor (EGFR) knockdown were used to assess ATBC-induced cellular phenotypes and associated signaling changes. Immune infiltration analysis was performed using OS transcriptomic datasets. Results A total of 226 overlapping targets between ATBC and OS were identified, with EGFR emerging as the topological hub. Molecular dynamics simulations suggested a stable predicted binding mode between ATBC and EGFR. ATBC promoted OS cell proliferation, migration, and invasion under the experimental conditions, accompanied by increased EGFR expression and AKT phosphorylation. EGFR knockdown attenuated the ATBC-induced cellular phenotypes. Immune infiltration analysis revealed exploratory associations between EGFR-co-expressed genes and distinct immune cell populations. Discussion These findings suggest that EGFR contributes to the pro-tumorigenic effects of ATBC in OS cells and indicate a potential link between EGFR-related networks and immune microenvironment remodeling. Further studies are warranted to clarify the underlying mechanisms and toxicological relevance of these observations.