4-Nonylphenol mediated tumor progression and immune modulation through CDK2: Computational and experimental validation.

As an endocrine-disrupting chemical, 4-nonylphenol (4-NP) has been found above safe limits in global water systems and associated with cancer progression. This multidisciplinary study systematically investigated the carcinogenic mechanisms of 4-NP by integrating network toxicology, machine learning, molecular docking simulations, single-cell RNA sequencing, and spatial transcriptomics. We identified oncogenic targets associated with 4-NP and performed pan-cancer expression profiling and prognostic analysis of 27 core genes, followed by machine learning algorithms to quantify the prognostic weights of these genes in different malignancies. Molecular docking identified six high-affinity 4-NP-binding targets (TNF, CDK2, PRKCA, ERBB2, CASP8, MAPK3); binding energies less than -7.0 (kcal/mol). Mechanistically, ERBB2, CDK2 and MAPK3 may be involved in the activation of the MAPK signaling pathway, and PRKCA plays a key regulatory role in this process, thus promoting oncogenic signaling. Meanwhile, TNF and CASP8 may lead to CD8⁺ T-cell exhaustion, thereby promoting the establishment of an immunosuppressive tumor microenvironment. Finally, we designed a 4-NP intervention model with concentrations ranging from 0.1 to 10 μmol/L, based on the upper concentration limits of polluted surface water and extreme upper values in heavily pollution, and validated its effect on promoting cell proliferation and CDK2 expression in two breast cancer cell lines. Our findings not only elucidate the potential role of 4-NP in tumorigenesis, but also establish a computational-experimental framework for environmental toxicant assessment. This work provides important insights for developing preventive strategies against pollutant-driven malignancies, bridging molecular toxicology and clinical cancer biology.