Identification of EGFR, MMP9, and AGTR1 as actionable therapeutic targets in acetyl tributyl citrate-driven breast cancer: integrated network toxicology and in vitro validation

Abstract

Background Growing exposure to acetyl tributyl citrate (ATBC) drives breast cancer (BC) progression and poses significant challenges to clinical management. However, the precise molecular targets that mediate ATBC-induced tumor progression remain largely elusive. Identifying these cell surface and intracellular targets is critical for the development of rational, integrated targeted therapeutic strategies. Methods A comprehensive Network Toxicology approach combined with multi-omics bioinformatics was employed to elucidate the molecular targets of ATBC in BC. Core hub genes were identified by intersecting potential ATBC targets with BC-related differentially expressed genes (DEGs). The diagnostic and immunological significance of these targets was evaluated. Finally, molecular docking and in vitro functional assays on BC cell lines were performed to validate target engagement and therapeutic potential. Results ATBC exposure demonstrated potential carcinogenic risk and promoted BC progression. We identified three critical hub targets: EGFR, MMP9, and AGTR1. Molecular docking confirmed high binding affinities between ATBC and these target proteins. In vitro validation revealed that ATBC significantly promoted cell proliferation and migration, and suppressed apoptosis in ER-positive MCF-7 cells, a mechanism driven by profound upregulation of EGFR, MMP9, and AGTR1 mRNA expression. Furthermore, the expression of these targets correlated significantly with immune cell infiltration within the tumor microenvironment. Conclusion ATBC may promote BC progression through upregulation of EGFR, MMP9, and AGTR1. These genes represent both potential diagnostic biomarkers and actionable therapeutic targets, providing a rational basis for repurposing existing EGFR inhibitors and angiotensin receptor blockers to treat environmentally driven BC.