Long-chain PFAS exposure and breast cancer-related molecular perturbations: An integrative analysis of NHANES and multi-omics data

Per- and polyfluoroalkyl substances (PFAS), especially perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS), are persistent organic pollutants associated with multiple adverse health outcomes, and growing evidence suggests a potential link to breast cancer. However, the molecular mechanisms underlying PFAS-associated breast carcinogenesis remain poorly defined. In this study, we integrated population-based epidemiological data, computational toxicology, transcriptomics, molecular docking, and cell-based assays to systematically elucidate the molecular pathways and toxic targets linking PFAS exposure to breast cancer-related molecular perturbations. A total of 4946 adult women from the NHANES 2005–2018 cohort were included. Weighted logistic regression and Bayesian kernel machine regression (BKMR) models demonstrated significant positive associations between serum PFOA/PFOS concentrations and breast cancer risk. Integrative network analysis identified 310 candidate PFAS-breast cancer targets, further refined to 33 top-ranked key genes enriched in oncogenic and immune-related signaling pathways. Transcriptomic discovery of PFAS-exposed breast cancer cells, cross model transcriptomic support, and cell-based assays highlighted CDK1, CDK2, and TP53 as candidate PFAS-responsive regulators. Molecular docking analysis predicted energetically favorable binding poses between PFOA/PFOS and these proteins (binding energies ≤ −6 kcal/mol). Cell-based assays further corroborated the upregulation of the core targets and provided preliminary evidence consistent with PFAS-associated direct or indirect target engagement. Collectively, these findings suggest that long-chain PFAS exposure may be linked to perturbation of cell-cycle control (via CDK1/CDK2) and stress-response/genomic-stability pathways (via TP53) within a broader oncogenic, endocrine-related, and immune-signaling network relevant to breast cancer.