Around 30% of patients with hormone receptor-positive (HR+) breast cancer acquire resistance to endocrine therapy combined with cyclin-dependent kinase 4/6 inhibitors (CDK4/6i), which are first-line treatments in metastatic settings. Therefore, we aimed to identify loci associated with resistance to endocrine therapy and CDK4/6i; this was achieved using retroviral vectors, which randomly insert gene-disrupting elements into the genome, causing gene expression alterations and potentially leading to therapy resistance. ER-positive ZR75.1 breast cancer cells transduced with retroviral vectors were treated with endocrine (tamoxifen, fulvestrant) or CDK4/6i monotherapies (abemaciclib, palbociclib, ribociclib) or a combination of fulvestrant and ribociclib. DNA was extracted, and virus integration sites (VISs) were characterized according to the detection frequency and read depth using next-generation sequencing (VIS-NGS). Resistance-associated VIS loci were identified when differentially presented in treated samples compared to controls. Well-established tamoxifen resistance genes (BCAR1, BCAR3, EGFR) were detected, enabling the validation of our approach. Thirty-seven VIS loci were associated with resistance to fulvestrant and ribociclib monotherapies. Twenty of these loci were also identified as candidates for resistance to other CDK4/6i and to fulvestrant and ribociclib combination therapy, including TRPS1 and TRIM24-genes that are involved in resistance to endocrine therapy but have not yet been associated with resistance to CDK4/6i. The identification of unique and shared resistance-associated loci highlights the complexity of resistance pathways.

Human immunodeficiency virus (HIV) infection remains a major global health burden. Untreated HIV infection leads to CD4+ T-cell depletion and severe immune dysfunction, resulting in opportunistic infections, neoplastic changes, and death. Highly active antiretroviral therapy (HAART) is currently the standard treatment for HIV infection, but it cannot eliminate latent reservoirs. Post-translational modifications (PTMs) regulate protein trafficking, function, and degradation, and their in-depth investigation plays a crucial role in identifying novel biomarkers and therapeutic targets. PTMs exert a central regulatory role in HIV infection by both enhancing host restriction factors and contributing to latent infection. This dual role offers novel insights into potential therapeutic targets for activating latent viruses to make them visible to the immune system. This review highlights numerous PTMs associated with HIV infection, including acetylation, phosphorylation, palmitoylation, etc., and assesses their potential for curing HIV infection.

Quinoa (Chenopodium quinoa Willd.) is a promising nutrient-rich crop, but its cultivation is threatened by widespread antimony (Sb) contamination in soils. While melatonin (MT) is known to alleviate abiotic stresses, its role in Sb tolerance is poorly understood. We screened 14 quinoa varieties and identified two cultivars with contrasting Sb accumulation: a high-accumulator (CL2) and a low-accumulator (No. 1). MT application effectively mitigated Sb-induced phytotoxicity, reducing Sb accumulation in roots by 33.81% and 55.43% in CL2 and No. 1, respectively, with significant reductions also observed in the shoots. Transcriptomic analysis identified 699 common differentially expressed genes, among which the transcription factor CqBBX8 was prioritized for functional characterization due to its strong induction by Sb and suppression by MT. Heterologous overexpression of CqBBX8 in tobacco promoted Sb uptake by upregulating metal transporter genes and aggravated oxidative damage by repressing antioxidant enzymes and enhancing NtRbohE expression. Results confirmed that CqBBX8 binds to the promoters of two putative Sb transporter genes, CqNRAMP and CqZIP, and activates their expression. It also binds to the promoters of CqPOD and CqRbohE, thereby differentially regulating these redox-related genes. Our findings establish CqBBX8 as a pivotal positive regulator of Sb uptake and sensitivity and reveal that MT enhances tolerance primarily by suppressing the CqBBX8 pathway. This work provides a novel mechanistic framework and a genetic target for breeding low-Sb crops to ensure food safety in contaminated regions.

Autophagy inhibition represents a promising therapeutic approach for the management of various cancers including nonsmall cell lung cancer (NSCLC). We previously reported SBP-7455, a dual inhibitor of unc-51-like kinase 1 (ULK1) and its homologue ULK2 and described its effects on triple-negative breast cancer (TNBC) cells. Herein we report the design, synthesis, and characterization of SBP-5147 and SBP-7501, two new dual ULK1/2 inhibitors that are cytotoxic against NSCLC cells, inhibit autophagic flux in A549 cells, and present greater oral exposure than SBP-7455 at a lower dose. In addition, SBP-5147 effectively modulates autophagy and increases the expression of major histocompatibility complex (MHC) class I in NSCLC cells, which may support the rationale for ULK1/2 inhibition as a strategy to overcome resistance to immunotherapy. Together these data support the use of ULK inhibitors as part of a cancer treatment strategy, either as a single agent or in combination with current therapies.

Hepatocellular carcinoma (HCC) exhibits a gender disparity, with a 2- to fourfold higher incidence in men, as attributed by previous studies to estrogen-mediated hepatoprotection versus the tumor-promoting role of androgens. This study investigated the effects of these sex hormones-testosterone and β-estradiol-on Nrf2 expression and its relation to key factors in metastasis, apoptosis, and drug resistance in HepG2 HCC cell line. HepG2 cells were exposed to testosterone or β-estradiol at viability-equivalent doses. Oxidative stress was quantified by total oxidant status (TOS), malondialdehyde (MDA), and total antioxidant capacity (TAC) assays. Apoptosis was evaluated by Annexin V/PI flow cytometry. mRNA levels of Nrf2, MRP-1 (multidrug resistance protein 1), MMP-9 (matrix metalloproteinase-9), BCL2, and SIRT1 were assessed via qRT-PCR. At viability-equivalent doses, β-estradiol elicited dose-dependent apoptosis in HepG2 cells (total: 47.4% at V₅₀ vs. 32.7% for testosterone; p < 0.001), predominantly late-stage (34.5% vs. 21.8%; p < 0.0001). Concomitantly, β-estradiol decreased TOS (maximal at V₄₀, p < 0.05) and MDA (all doses, p < 0.05) while increasing TAC (medium/high doses, p < 0.05)-effects not observed with testosterone (p > 0.05). β-estradiol induced downregulation of Nrf2, BCL2, MMP-9, and MRP-1 mRNA (moderate/high doses; p < 0.05 vs. control). Conversely, testosterone upregulated SIRT1 across doses (p < 0.05), unaffected by β-estradiol (p > 0.05). β-Estradiol mitigates oxidative stress, induces apoptosis, and suppresses pro-survival, metastatic, and chemoresistant pathways-contrasting testosterone's minimal effects. These findings align with HCC's male predominance and highlight the hormone-modulated strategies' potential for future therapeutic exploration.

The biological aging of vascular smooth muscle cells (VSMCs) is a critical event contributing to vascular aging and age-related diseases. The objective of this study is to assess the potential mechanism by which eugenol inhibits vascular senescence by downregulating milk fat globule-EGF factor 8 (MFG-E8) expression in VSMCs and vascular tissues. A model of angiotensin II (Ang II)-induced vascular aging in mice and a premature aging model in human vascular smooth muscle cells (HVSMCs) were established to assess the efficacy of eugenol intervention, with valsartan, an Ang II receptor antagonist, serving as a positive control drug. Senescence-associated-β-galactosidase (SA-β-gal) staining, along with the detection of senescence marker molecules p21 and p53, were used to assess the senescence status of cells and vascular tissues. The expression level of MFG-E8 was detected using immunohistochemistry, reverse transcription-quantitative PCR, and western blot analysis. HVSMC cell lines with MFG-E8 knockdown and overexpression were generated using short hairpin RNA and plasmid overexpression methods, respectively, to assess the role of MFG-E8 in the anti-senescence effects of eugenol on VSMCs. Eugenol inhibited Ang II-induced premature senescence in both vascular tissues and HVSMCs, significantly enhancing arterial stiffness and structural changes in aged vessels. It also attenuated the senescence-associated secretory phenotype (SASP) and enhanced the proliferative activity of senescent VSMCs. Ang II exposure led to increased MFG-E8 expression in cells and vascular tissues, a change that eugenol was able to reverse. Knockdown of MFG-E8 suppressed the Ang II-induced senescence phenotype in HVSMCs, whereas overexpression of MFG-E8 directly induced HVSMC senescence and counteracted the anti-aging effects of eugenol. Eugenol prevents Ang II-induced aging in VSMCs and vascular tissues by downregulating MFG-E8 expression, underscoring its potential as an antiaging drug.

Grass carp (Ctenopharyngodon idellus), China's most valuable freshwater aquaculture species, exhibits growth and nutrient utilization efficiency that are highly dependent on feed quality. After macronutrient balancing, nano-selenium (nano-Se) supplementation becomes critical for enhancing health and profitability. Although nano-Se has been observed to alleviate oxidative stress and inflammation, the molecular mechanisms underlying its hepatoprotective effects following long-term administration remain systematically uncharacterized. To fill this gap, this study utilized transcriptomic and metabolomic technologies to investigate the beneficial alterations in the liver of grass carp following 30 weeks of nano-Se feeding. Ninety juvenile grass carp were randomly allocated to either a control group (basal diet) or a nano-Se group (basal diet + 0.6 mg/kg nano-Se); livers were harvested for omics analyses at the end of the 30-week feeding period. Transcriptomic analysis initially identified 533 differentially expressed genes (110 up-regulated, 423 down-regulated). Gene Ontology (GO) functional enrichment analysis indicated that these genes were primarily involved in biological processes such as metabolic processes, biological regulation, and stress response, suggesting that nano-Se broadly regulates hepatic metabolic activity and stress adaptability. Further Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed significant enrichment in the "protein digestion and absorption" pathway. Key genes in this pathway, including collagen VI α1/2 (COL6A1/2), elastase ELA2/3 L, and amino acid transporters SLC3A1 and SLC7A9, were significantly down-regulated, indicating that nano-Se may mitigate oxidative stress-induced micro-damage in hepatocytes, thereby reducing the liver's demand for damage repair and fibrotic processes. Metabolomic analysis detected 1404 metabolites, with 267 differentially metabolites (198 up-regulated, 69 down-regulated) spanning multiple metabolic categories such as amino acids, lipids, and cofactors. Glycerophospholipids (GP) and sphingolipids (SP) were significantly up-regulated, while bile acid metabolites were down-regulated. Related metabolic pathway analysis showed that "glycerophospholipid metabolism" and "linoleic acid metabolism" pathways were significantly activated. Glycerophospholipids and sphingolipids are major components of cell membranes; their increased levels may enhance the integrity and stability of hepatocyte membranes. Concurrently, enhanced linoleic acid metabolism may contribute to energy supply and inflammation regulation. These changes collectively suggest that nano-Se may improve hepatic redox homeostasis and metabolic balance by remodeling hepatocyte membrane lipid composition and optimizing energy metabolism pathways. Collectively, nano-Se alleviates oxidative injury and maintains metabolic homeostasis in the grass-carp liver through the coordinated modulation of amino-acid, lipid and immune-related pathways, thereby providing a theoretical basis for its long-term, safe application in aquafeeds; future work is still required to verify these findings by measuring antioxidant-enzyme activities and to optimize dosage through graded-dose experiments.

Waterlogging stress typically causes submergence or partial submergence stress in plants, which negatively impacts agricultural production, from seed germination to vegetative and reproductive growth. Uniconazole (S3307) was recently used to minimize the damage caused by waterlogging. Here, we investigate the effects of S3307 on the growth, development, and yield of soybean under waterlogging stress. Morphological and physiological indexes, as well as the transcriptome and metabolome of soybean were analyzed in control condition and waterlogged condition with (WS) or without (W) spraying S3307. The results showed that waterlogging stress led to growth inhibition and reduced activity of antioxidant enzymes in soybean plants, resulting in a large accumulation of MDA content and ultimately causing yield reduction. Foliar spraying of S3307 increased stem diameter, reduced plant height, increased dry matter accumulation, increased antioxidant enzyme activities, inhibited MDA accumulation, and alleviated the yield reduction. The combined transcriptomics and metabolomics investigation demonstrated that, compared to Control, both W and WS treatments generated similar directional changes in triterpenoid metabolism and stress response pathways, suggesting that waterlogging stress activated these defense-related pathways. Notably, the strength of these reactions was considerably greater in the WS treatment than in the W treatment. Under waterlogging stress, foliar application of S3307 increased the expression of genes involved in flavonoid production and increased the accumulation of unsaturated fatty acids-substances known to help in cell membrane stability-when compared to the W treatment. In summary, our findings show that S3307 does not alter the direction of waterlogging-induced reactions but rather greatly increases their strength, thereby positively regulating soybean waterlogging tolerance. This establishes a biochemical mechanism and theoretical foundation for using plant growth regulators to reduce stress.

Neuroblastoma, originating from the sympathetic neural crest, is the most prevalent extracranial solid tumor in children. Amplification of MYCN is a widely recognized indicator of poor prognosis in neuroblastoma. However, the structural properties of the N-Myc protein encoded by MYCN have impeded the development of direct inhibitors with favorable drug-like properties. This study aimed to investigate the upstream regulatory mechanisms of N-Myc stabilization in neuroblastoma and explore potential therapeutic strategies targeting these mechanisms.

NF2-related schwannomatosis (NF2-SWN) is a debilitating condition, characterized by bilateral vestibular schwannomas (VSs) that progressively cause irreversible sensorineural hearing loss. Current management relies on surgery or radiotherapy, while bevacizumab (αVEGF) is used off-label, with variable and often transient efficacy. Effective therapies that durably suppress tumor growth and preserve hearing are urgently needed. Although immune checkpoint inhibitors have transformed cancer treatment, their efficacy in non-malignant tumors such as VS remains unclear. Here, we evaluate combined anti-PD1 (αPD1) and αVEGF therapy in two syngeneic, immune-competent VS models. Combination treatment significantly outperforms either monotherapy, inhibiting tumor growth and preventing hearing loss. Mechanistically, αVEGF enhances αPD1 efficacy by normalizing tumor vasculature, improving drug delivery and immune cell infiltration, and promoting cytotoxicity of T and NK cells via NKG2D upregulation. Combined treatment effectively controls tumor growth that progresses despite anti-VEGF therapy. These findings support αPD1 and αVEGF combination therapy as a promising strategy for NF2-SWN.

Recent data suggests one fungus, Aspergillus fumigatus, causes more deaths annually than HIV or malaria combined. Coupled with rapid emergence of antifungal drug resistance, the limited range of effective treatments, and mortality rates of >50%, aspergillosis represents a major challenge in infectious diseases. Recent studies have identified long-noncoding RNAs (lncRNAs) involved in drug resistance and virulence in pathogenic yeasts such as Candida spp. However, there is very limited knowledge of lncRNAs in human pathogenic moulds, including A. fumigatus. Here we exploit transcriptomics data of A. fumigatus exposed to different environments to annotate transcripts mapping to 2388 genomic loci. After manual curation we generate a database of over 1000 lncRNAs. We observe that the lncRNAs display orchestrated transcriptional profiles upon drug treatment and many are proximal to genes involved in azole sensitivity. We knock out a set of intergenic lncRNAs and perform a large-scale phenotypic analysis to identify 60 lncRNA mutants displaying condition-dependent fitness changes with 35 mutants exhibiting a positive growth phenotype under azole stress. Overall, this study generates and experimentally validates an important resource that will enable the wider research community to increase understanding of the functional importance of lncRNAs in A. fumigatus, including their involvement in drug sensitivity.

Ferroptosis is a newly identified programmed cell death induced by iron-driven lipid peroxidation and implicated as a potential approach for tumor treatment. Breast tumors develop in a complex microenvironment whose main component is adipose tissue and gain aggressiveness through increased fatty acid uptake. Here, we demonstrated that palmitic acid (PA) induced ferroptosis in triple negative breast cancers (TNBC). We found that PA increases the protein expression levels of the long-chain fatty acid transporter CD36, leading to increased lipid uptake. Mechanistically, overexpression of CD36 increases lipid peroxidation, mitochondrial ROS production, the labile iron pool, and especially Fe2+ content. Additionally, we found increased expression of ferroptotic target genes (HMOX1, ACSL1, SAT1) and decreased of anti-ferroptotic genes (GPX4 and FSP1) in TNBC following PA exposure. Overexpression of CD36 did not induce ferroptosis in estrogen receptor positive breast cancer. Clinically, higher CD36 expression correlated with the luminal androgen receptor (LAR) subtype of TNBC, known to exhibit a higher sensitivity to ferroptosis. Altogether, these data provide evidence for an essential role of the CD36 protein in the ferroptotic process induced by the saturated fatty acid PA, opening potential new therapeutic approaches promoting ferroptosis in the most aggressive breast cancers.

Hyperuricemia (HUA) is a chronic metabolic disease which has been previously observed to be associated with cortisol metabolism disorders (pseudohypoadrenalism). In this study, we aimed to investigate the efficacy as well as mechanism of Eurycoma longifolia Jack (TkA) on alleviating cortisol metabolism. Oral administration of TkA significantly decreased serum uric acid levels and urinary cortisol in HUA mice. TkA improved HPA axis function and upregulated the levels of adrenal Hsd3b2, Cyp21a1 and Cyp11b1. In the liver, TkA upregulated the expression of Srd5a1 and Akr1c4, promoting the conversion from cortisol to 5α-tetrahydrocortisol (P < 0.001). TNFα was found to be the principal driver of reduced SRD5A1. By activating NF-κB pathway, recruited DNA methyltransferase (DNMT) binding with the CpG islands increased methylation level of Srd5a1. Our findings highlight that eurycomanol significantly inhibited the activation of IKKβ/IκBα/NF-κB/DNMT pathway as well as up-regulated hepatic SRD5A1, thereby restoring the systemic cortisol metabolic homeostasis under HUA.

The multinucleated syncytiotrophoblast (STB) at the maternal-fetal interface is formed through the continuous fusion of mononucleated cytotrophoblasts (CTBs). Estradiol and protein glycosylation are known to participate in trophoblast syncytialization. O-fucosyltransferase 2 (poFUT2), which catalyzes protein O-fucosylation, has been implicated in placental development. However, the exact role of poFUT2 in trophoblast syncytialization remains unclear. The aim of the present study is to investigate the function of the estradiol-poFUT2 axis in trophoblast syncytialization. Here, by applying immunohistochemistry, we found that poFUT2 expression was decreased in syncytiotrophoblast of placental tissues from preeclampsia patients. Employing a cell-model to induce trophoblast syncytialization in vitro, we demonstrated that poFUT2 promotes trophoblast cell fusion. Mechanistically, during pregnancy, elevated estradiol upregulated the expression of poFUT2 and enhanced syncytium formation. Chromatin immunoprecipitation and co-immunoprecipitation assays indicated that the regulation depends on the participation of SP1 in the estrogen receptor 2 (ESR2)-mediated regulation of the poFUT2 gene. Conclusively, our findings demonstrated that estradiol upregulated poFUT2 expression via the ESR2/SP1 complex, increasing trophoblast cell differentiation and fusion.

EGFR-TKI-resistant cell lines were established by long-term exposure to gefitinib, afatinib, and osimertinib via the PC9 model. This model helps study EGFR-TKI resistance mechanisms in non-small cell lung cancer (NSCLC) and may offer insights to improve treatment outcomes, particularly for patients unresponsive to anti-PD-1/PD-L1 therapies. We investigated molecular alterations in these resistant cell lines using multi-omics techniques, including genome sequencing, proteomics, and transcriptomics. Differential dependencies on EGFR downstream pathways were observed among resistant cell lines. Immune evasion mechanisms were analyzed to identify alterations in EGFR downstream pathways and unfolded protein response (UPR) elements contributing to immune-related transcriptional patterns. Afatinib-resistant cells exhibited the most pronounced immunosuppressive-like transcriptional features compared to gefitinib- and osimertinib-resistant cells. This was characterized by elevated PD-L1 expression, minimal changes in MHC class 1 levels, down-regulation of shared immune-modulated gene sets, and a cytokine profile favoring immunosuppression (e.g., lower IL-6, CXCL10, IFN-γ, CCL22; higher IL-8). These findings suggest a poor prognosis and limited efficacy of anti-PD-1/PD-L1 immunotherapy in NSCLC patients with similar resistance profiles. Notably, inhibition of IRE1α signaling partially reversed this immunosuppressive-like transcriptional signature, indicating a potential strategy to restore immune responsiveness in afatinib-resistant NSCLC. Our study underscores the distinct remodeling effects of different EGFR-TKIs on tumor-cell-intrinsic immunoregulatory pathways. Targeting endoplasmic reticulum (ER) stress pathways alongside immune checkpoint inhibitors may be crucial for overcoming resistance mechanisms identified here. These insights provide a rationale for personalized treatment strategies tailored to the immune-related gene-expression profiles observed in EGFR-TKI-resistant NSCLC models, aiming to enhance therapeutic responses and improve clinical outcomes.

RKIP (Raf Kinaes Inhibitory Protein) is ubiquitously expressed in almost all normal tissues of metazoans. Consistent with its negative regulatory role in cell proliferation and survival, RKIP expression is progressively downregulated in cancer accompanied with a worse prognosis and disease outcome. Experiments with cancer cell lines and genetically engineered mice have demonstrated that the expression level of RKIP is a driving factor in determining the disease outcome of cancer, and that restoring RKIP expression is a promising option for therapeutic treatment of low-RKIP-expressing cancers. RKIP expression is mainly regulated at the transcription level such that its expression is tuned down, but not shut off, and is poised to be reactivated. In this study we developed a RKIP gene promoter knock-in reporter breast cancer cell line to screen for compounds that can regulate RKIP transcription in cancer. Using this modified cell line, we were able to identify eight FDA approved compounds that increase RKIP promoter activity by a minimum of 2-fold and can potentially be re-purposed for therapeutic treatment of low-RKIP-expressing breast cancer.

Lung adenocarcinoma (LUAD) is a major lung cancer subtype influenced by environmental factors. Benzo[a]anthracene (BaA), a common Group 2B carcinogen found in pollutants, smoke, and food, shows genotoxic and oncogenic activity; however, its specific mechanisms in LUAD pathogenesis remain unclear and warrant systematic investigation.

Hepatoblastoma is the most common pediatric liver tumor, and it primarily affects young children. Despite improved treatment results due to modern chemotherapy and advanced surgical techniques, there is urgent need to improve the long-term prognosis of hepatoblastoma patients. Elevated expression of transcription factor GATA4 has been associated with pro-tumorigenic functions in hepatoblastoma. Herein, we explored the effects of a novel series of GATA4-targeted compounds (3i-2010-3i-2014) in several hepatoblastoma cell models. To this end, we assessed cell viability (2D and 3D), caspase 3/7 activity, cell cycle process with automated fluorescence microscopy, and RNA and protein expression using quantitative PCR, RNA-sequencing, and western blotting after treating cells with GATA4-targeted compounds. The GATA4-targeted compounds reduced hepatoblastoma cell viability but affected also healthy control cells when high doses were applied. Out of the five compounds used, the 3i-2012 was the most potent. This compound induced G0/G1 phase cell cycle arrest, increased caspase activity, and resulted in changes in transcriptome of hepatoblastoma cells. The differentially expressed genes (DEGs) were associated with cell cycle regulation. Transcription factor target analysis demonstrated that a notable proportion of DEGs were likely regulated by GATA4. In conclusion, 3i-2012 decreases hepatoblastoma cell survival by disturbing cell cycle regulation in the tumor cells.

Oral squamous cell carcinoma (OSCC) is a prevalent and aggressive malignancy with a persistently high mortality rate, largely attributable to therapy resistance and tumor recurrence. This review comprehensively explores the critical interplay between epigenetic dysregulation and the tumor microenvironment (TME) in driving OSCC progression. We detail how key epigenetic mechanisms, including DNA methylation, histone modifications, and non-coding RNAs (ncRNAs), intrinsically transform cancer cells and actively orchestrate pro-tumorigenic TME. These alterations substantially contribute to resistance against conventional therapies. Furthermore, we discuss the therapeutic potential of targeting these pathways using epigenetic drugs (epi-drugs), such as DNA methyltransferase (DNMT) inhibitors and histone deacetylase (HDAC) inhibitors, as well as engineered extracellular vesicles (EVs). The primary objective of this review is to synthesize current knowledge on the epigenetic-TME axis, thereby providing a mechanistic foundation for developing novel therapeutic strategies. We emphasize that rational combinations of epigenetic-targeting agents with conventional treatments or immunotherapy hold significant promise for overcoming drug resistance and improving clinical outcomes in OSCC patients.

Diethyl phthalate (DEP) is a ubiquitous environmental endocrine-disrupting chemical (EDC). Epidemiological studies have suggested a potential association between DEP exposure and an increased risk of endometrial cancer (EC); however, its underlying molecular mechanisms remain largely unclear. Four GEO datasets were integrated, and differential expression analysis combined with weighted gene co-expression network analysis (WGCNA) was performed to identify candidate genes potentially linking DEP exposure to EC. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to explore relevant signaling pathways. Machine learning models, coupled with Shapley Additive Explanations (SHAP), were employed to prioritize key genes. Molecular docking and molecular dynamics (MD) simulations were used to assess the binding affinity between DEP and the identified targets. A series of in vitro experiments in EC cell lines were subsequently conducted to validate the biological effects of DEP. Nineteen overlapping DEP-EC genes were identified, predominantly enriched in the MAPK, cAMP, and cGMP-PKG signaling pathways. Among them, FOS, NR4A1, ADRA2C, JUN, and SLC6A2 were prioritized as core genes through machine learning and SHAP analysis. Molecular simulations confirmed stable binding between DEP and these targets. In vitro assays demonstrated that DEP exposure induces oxidative stress, significantly enhances ERK1/2 and AKT phosphorylation, upregulates Cyclin D1/CDK4 expression, promotes G1/S phase transition, and facilitates EC cell proliferation. These findings suggest that DEP may promote endometrial carcinogenesis by triggering oxidative stress-mediated signaling crosstalk and accelerating cell cycle progression. This study establishes a multi-layered methodological framework-from computational screening and machine learning to experimental validation-offering novel mechanistic insights into the carcinogenic potential of environmental endocrine disruptors such as DEP.