Di-(2-ethylhexyl) terephthalate promotes breast cancer progression: Multi-omics integrated experimental validation

Di-(2-ethylhexyl) terephthalate (DOTP), as an alternative to phthalate plasticizers, has been widely used in sensitive fields such as food packaging and medical devices. However, there remains a gap in the assessment of its systemic carcinogenic risk. This study systematically analyzed the potential carcinogenic mechanisms of DOTP by integrating network toxicology methods such as gene enrichment analysis, machine learning algorithms, molecular docking, single-cell sequencing, and spatial transcriptomics. We screened 24 core carcinogenic genes of DOTP using 11 topological algorithms, conducted cross-cancer expression and prognostic analyses, and quantified the prognostic risk weights of these genes in tumors using machine learning algorithms. Subsequently, we used molecular docking to identify six carcinogenic targets with high affinity for DOTP (PIK3CA, PTPN11, ESR1, PPARG, PTGS2, and MAPK1). Finally, using colony formation assays and Western blotting analysis, we confirmed that DOTP exposure can promote the proliferation of tumor cells. Both this proliferative effect and the induced upregulation of PTPN11 and ESR1 protein levels exhibited concentration-dependent enhancement. The study suggests that DOTP may exhibit carcinogenic potential by interfering with cell proliferation, gene transcription regulation, the tumor microenvironment, and related carcinogenic signaling pathways (primarily the PI3K-Akt-mTOR pathway). This study reveals the molecular mechanisms of DOTP in the occurrence and development of common tumors, providing a new theoretical framework for studying the carcinogenic mechanisms of environmental pollutants and laying the foundation for targeted prevention.