Food and Chemical Toxicology

BPA exposure enhances the metastatic aggression of ovarian cancer through the ERα/AKT/mTOR/HIF-1α signaling axis

Long-term exposure to bisphenol A (BPA) in humans may promote ovarian cancer development. In present study, the mechanisms by which BPA mediates the aggression metastatic behavior of ovarian cancer were investigated in vitro/in vivo. The results showed that BPA (10 μM) significantly promoted the proliferation, migration and invasion of human ovarian cancer cells (ES-2 and OVCAR-3 cells); moreover, it promoted ES-2 and OVCAR-3 cell glucose uptake, lactic acid release and intracellular ATP synthesis. After administration of 5 μg/kg/day BPA, tumor volume was increased compared with that in control group. KEGG and GO enrichment analyses showed that the genes from ES-2 cell in 10 μM BPA-treated group were enriched mainly in central carbon metabolism and PI3K-AKT signaling pathway. Then, qRT‒PCR and western blotting results showed that BPA (10 μM) increased the mRNA and protein expression levels of glycolysis-related genes and mTOR, p-AKT HIF-1α and ERα in vitro/vivo; whereas this effect was reduced after treatment with the ERα inhibitor methyl-piperidino-pyrazole. Furthermore, coimmunoprecipitation and mass spectrometry showed that BPA promoted the direct interaction of ERα with lactate dehydrogenase A. These results show that BPA directly promoted the proliferation, migration and invasion of ovarian cancer cells through the ERα/AKT/mTOR/HIF-1α signaling axis to enhance glycolysis.

CDDO-Me triggers ROS-dependent ferroptosis and apoptosis in cervical cancer via targeting PI3K/Nrf2 pathway

Cervical cancer (CC) ranks as one of the most common types of malignant tumors affecting women. CDDO-Me is derived from oleanolic acid, a pentacyclic triterpenoid obtained by chemical structural modification, which has been shown anti-tumor effects. CC cell proliferation was decreased by CDDO-Me both in vitro and in vivo. Mechanistically, CDDO-Me led to a reduction of intracellular mitochondrial volume and crista, or an increase of membrane density. The levels of ROS, Fe CDDO-Me induces CC cell lines apoptosis and ferroptosis through the PI3K/Nrf2 signaling pathway, for exerting anti-proliferation effects.

Inhibitory effect and mechanism of purified saponin from Polygonatum cyrtonema Hua on human ovarian cancer cell A2780

Polygonatum cyrtonema Hua (PcH) is a traditional medicinal and edible homologous plant. Here, the anticancer activity of different purified PcH saponins (PcHs) on the ovarian cancer (OC) cell A2780 were investigated. The PcHs were purified by AB-8. Their effects on the apoptosis, migration and invasion of A2780 were explored by CCK8, wound healing assay, Hoechst 33258 staining, Annexin V-FITC/PI and intracellular reactive oxygen species (ROS) measurement. Moreover, western blot and realtime-PCR confirmation were performed for the associated protein and mRNA targets. The results showed that the content of PcHs increased from 0.31 ± 0.02 mg/mL to 0.79 ± 0.02 mg/mL after purification. The inhibition rate of purified PcHs on A2780 could reach 66.76 ± 0.56%. Specifically, it promoted cell apoptosis, disturbed the cell cycle, increased the ROS levels, regulated the expression levels of pro-apoptotic and anti-apoptotic proteins. PcHs can also regulate the expression level of proteins related to the invasion and migration characteristics of A2780, the mRNA expression level related to the P13K-Akt signal transduction process, and the extracellular matrix-receptor interaction pathway, as well as drug resistance. Overall, this study provides a new idea for the treatment of OC.

Aspartame promotes ovarian cancer progression: A multi-omics study integrating mendelian randomization, network toxicology, and in vitro validation

The causal relationship between the artificial sweetener aspartame and ovarian cancer (OC), a highly lethal malignancy, remains unclear. This study, therefore, employed a multi-omics approach to investigate this causal link and its potential mechanisms. First, a Mendelian Randomization (MR) analysis indicated that genetically predicted aspartame intake is associated with an increased risk of OC (OR = 2.10, 95 % CI: 1.06-4.18). Next, by integrating network toxicology with machine learning algorithms (LASSO, SVM, and Random Forest), we identified AURKA, CCND1, and RAD51 as potential core target genes. Further validation using multi-omics data from bulk and single-cell RNA sequencing confirmed that these three genes are upregulated in OC tissues. A subsequent MR analysis also provided causal evidence that high expression of CCND1 increases the risk of OC. Furthermore, molecular docking simulations showed that aspartame could form stable bonds with all three target proteins. Finally, in vitro experiments demonstrated that aspartame significantly promoted the malignant phenotypes of OC cells and regulated the expression of these core genes. In conclusion, this study suggests that aspartame may promote ovarian cancer development, potentially by upregulating the expression of key genes such as AURKA, CCND1, and RAD51. These findings provide new evidence for evaluating the safety of aspartame.

Zearalenone promotes endometrial cancer cell migration and invasion via activation of estrogen receptor-mediated Rho/ROCK/PMLC signaling pathway

Zearalenone (ZEA), has emerged as a potential endocrine-disrupting chemical (EDC). Previous results show ZEA effects on endometrial stromal cell apoptosis, migration, and growth of endometriosis. Despite the reported presence of ZEA in Endometrial Cancer (EC) patient's blood and tissues, ZEA-induced EC promotion and its mechanism/s remain elusive. In this study, Ishikawa cells were used to investigate the ZEA effects on Ishikawa cell migration, invasion, and the underlying mechanism involved in these events. Ishikawa cells were exposed to low concentrations of ZEA (5, 25, and 125 nM) for 48 h, and morphological alterations, migration, invasion, markers associated with epithelial-mesenchymal transition (EMT), E-cadherin, Vimentin, RhoA/ROCK/PMLC pathway activation were analyzed. ZEA (25 nM) exposure caused morphological alterations like stress fiber, filopodia formation, loss of cell adhesion, and a significant increase in migration and invasive potential in extracellular matrix-coated porous membranes. Moreover, ZEA exposure also increases the Rho-GTPase activity and expression of pathway mediators, GEFH1, RhoA, ROCK1+2, CDC42, and PMLC/MLC. Furthermore, pre-treatment with specific pharmacological inhibitors for Estrogen receptor-alpha (ER-α) and ROCK attenuate the ZEA-induced stress fiber formation and altered expression of E-cadherin, Vimentin, and Rho/ROCK/PMLC pathway mediators. These findings suggest that Rho/ROCK/PMLC signaling pathways are involved in ZEA-induced Ishikawa cell migration and invasion.

Bisphenol A exposure modulates ovarian cancer gene expression and oxidative stress markers: a case-control study

In this case-control study conducted at Cairo's National Cancer Institute, the association between bisphenol A (BPA), an endocrine-disrupting chemical (EDC), and ovarian cancer was investigated. BPA levels in the urine, oxidative stress markers (reactive oxygen species (ROS) and superoxide dismutase (SOD) activity), and Keratin 4 (KRT4) gene expression were analyzed in 30 patients and 30 controls. Significant risk factors for BPA exposure included consuming microwave meals, consuming canned beverages, using polyvinyl chloride (PVC) food storage, eating fast food, handling thermal paper, exposure to dust, and recurrent hospitalizations. Compared with normal controls, ovarian cancer patients presented increased BPA levels, ROS, and KRT4 expression, along with reduced SOD activity (p < 0.001). A strong positive correlation was found between BPA and KRT4, indicating that KRT4 may be a potential biomarker. The cutoff values for urinary BPA and KRT4 achieved 100 % sensitivity and specificity in distinguishing patients from controls. These findings suggest that BPA plays a role in ovarian cancer pathogenesis, likely through oxidative stress and gene dysregulation. This study emphasizes the importance of minimizing BPA exposure (e.g., by reducing the use of canned or packaged foods) and calls for larger studies to further investigate the role of EDCs in hormone-dependent cancers.

Targeting the PERK/NRF2 pathway: Enhancing cisplatin efficacy in resistant ovarian cancer cells via MRP1 and ROS modulation

Overcoming chemotherapy resistance remains a pivotal challenge in ovarian cancer treatment. This study investigates the potential of combining cisplatin with GSK2606414, a PERK inhibitor, to enhance therapeutic efficacy against cisplatin-resistant ovarian cancer cells. cisplatin resistance is driven by the PERK/NRF2 pathway, which activates MRP1 upregulation and antioxidant defenses, thus promoting cell survival. By inhibiting PERK phosphorylation, GSK2606414 disrupts this pathway, downregulating MRP1 and increasing oxidative stress to sensitize cancer cells to cisplatin. Our findings reveal that the GSK2606414-cisplatin combination significantly reduces cell viability and proliferation, particularly in resistant cell lines, allowing for dose reduction and potentially lower side effects. The combined therapy also amplifies, increasing Caspase-12 and CHOP protein levels during endoplasmic reticulum stress. By targeting the p-PERK/p-NRF2/MRP1 and p-PERK/p-NRF2/SOD pathways, GSK2606414 decreases MRP1 expression and elevates ROS levels, rendering resistant cells more susceptible to chemotherapy. Additionally, this combination boosts intracellular cisplatin accumulation in both cisplatin-sensitive and -resistant ovarian cancer cell lines, reinforcing its cytotoxic impact. These findings underscore the promise of GSK2606414 and cisplatin co-treatment as a potent strategy to counteract ovarian cancer resistance. This combination could potentially advance therapeutic outcomes and provide a new pharmacological approach to resistant cancers.

Evaluation of effects of bisphenol analogs AF, S, and F on viability, proliferation, production of selected cancer-related factors, and expression of selected transcripts in Caov-3 human ovarian epithelial cell line

Bisphenol A (BPA) has been a substantial additive in plastics until the reports on its adverse effects have led to its restrictions and replacement. Monitoring studies document the increasing occurrence of bisphenol analogs, however, data on their effects and risks is still insufficient. Based on the indications that BPA might contribute to ovarian cancer pathogenesis, we examined effects of the analogs AF (BPAF), S (BPS) and F (BPF) (10

6PPD-quinone promotes ovarian cancer progression: Insights from network toxicology, machine learning, and in vitro validation

6PPD-quinone (6PPD-Q), a toxic tire-derived antioxidant transformation product, is a pervasive environmental contaminant linked to reproductive toxicity. However, its role in ovarian cancer (OC) remains elusive. We integrated network toxicology with transcriptomic analysis to elucidate the oncogenic mechanisms of 6PPD-Q. By screening the GEO, SwissTargetPrediction, and SEA databases, we identified 26 intersection targets. Utilizing machine learning and SHAP analysis, five core biomarkers-DDR1, ABL1, PDE2A, FRK, and F10-were prioritized. Molecular docking demonstrated high binding affinities between 6PPD-Q and these core proteins. In vitro validation, including CCK-8, plate colony formation, and qRT-PCR assays, confirmed that 6PPD-Q exposure significantly promotes OC cell proliferation. Mechanistically, 6PPD-Q may regulate the expression of hub targets to affect OC progression. This study establishes a definitive "exposure-target-phenotype" chain, characterizing 6PPD-Q as a potential environmental promoter of OC and suggest potential intervention targets.

Dibutyl phthalate (DBP) induces cervical injury and promotes malignant transformation of cervical epithelial cells

Dibutyl phthalate (DBP), a widespread phthalate with reproductive toxicity and potential carcinogenicity, its cervical effects remain unclear. This study explored DBP's cervical toxicity and mechanisms via epidemiological analysis, network toxicology, and in vivo/in vitro models. Urinary 6 phthalate metabolites in 104 cervical cancer (CC) patients and 104 controls (detected by UPLC-MS/MS) showed elevated levels of MBP, MEHP, MEOHP, MEHHP, MECPP, and ΣDEHP in CC patients; MBP had the strongest CC risk association (adjusted OR = 2.54, P < 0.001). Network toxicology identified 9 core targets of DBP (e.g., CASP3, MAPK8/14, ESR1) and key pathways involved (TNF, MAPK, apoptosis, oxidative stress, etc.). Short-term DBP exposure (mice: 10-50 mg/kg/day; HcerEpic cells: 100-400 μM) induced cervical injury/oxidative stress, suppressed NRF2, and activated MAPK/NF-κB; N-acetylcysteine (NAC) supplementation mitigated damages. Long-term exposure to environmentally relevant DBP concentrations (10

Osthole inhibits proliferation and induces autophagy-dependent ferroptosis of cervical cancer cells through induction of TFR1

Osthole (OST), a natural coumarin, exerts its anticancer effects by inducing cancer cell necroptosis, apoptosis, pyroptosis or autophagy. In He La cells, OST cause mitochondrial atrophy, increased membrane density and reduced cristae, which are characteristics of ferroptosis. However, the mechanisms of OST-induced ferroptosis in cervical cancer cells remain unclear. In the present study, we found that OST inhibited He La cells' proliferation which triggered ferroptosis. Further studies showed that ferroptosis or autophagy inhibition could reverse the inhibitory effects of OST on He La cell proliferation, and significantly inhibited ferroptosis and autophagy induced by OST. Moreover, autophagy stimulation enhanced the ferroptosis induction of OST, while ferroptosis induction in turn enhanced the autophagy levels caused by OST. Furthermore, Transferrin Receptor 1 (TRF-1) silencing effectively reduced OST-induced ferroptosis and autophagy, while TFR1 overexpression notably enhanced OST-induced ferroptosis and autophagy in He La cells. In addition, animal experiments demonstrated that overexpression of TFR1 effectively suppressed tumor growth in cervical cancer He La cells, and its expression was associated with the activation of ferroptosis and autophagy. In conclusion, our study revealed the detailed mechanism of OST-induced He La cells ferroptosis and autophagy, and the association between autophagy and ferroptosis in OST-induced cell death.

Piperlongumine induces apoptosis and autophagy via the PI3K/Akt/mTOR pathway in KB human cervical cancer cells

Natural products are continuously being researched to develop safe and effective treatment options for cervical cancer, the fourth most common cancer in women. Piperlongumine (PL), an amide alkaloid mainly present in long pepper, exhibits neuroprotective and anti-cancer properties. However, the specific effect of PL in cervical cancer and the relationship between the anti-cancer pathway and autophagy remain unclear. Therefore, we aimed to investigate PL-induced apoptosis in KB human cervical cancer cells and the relationship between apoptosis and autophagy therein. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and wound-healing assays showed that PL treatment suppressed KB cell viability and proliferation. Apoptosis was identified through 4',6-diamidino-2-phenylindole and annexin V-propidium iodide staining, increased cleaved-poly (ADP-ribose) polymerase and Bcl-2 associated X levels, and decreased B cell lymphoma 2 levels. Acridine orange staining and increased microtubule-associated protein 1A/1B-light chain 3-II and Beclin-1 levels confirmed autophagy. We determined that KB cell-related autophagy exerted cytoprotective effects using the autophagy inhibitors 3-methyladenine and hydroxychloroquine. PL treatment promoted apoptosis by inhibiting the phosphatidylinositol-3-kinase (PI3K)/protein kinase B/mammalian target of rapamycin pathway in KB cells; inhibiting the pathway using PI3K inhibitors increased autophagy. We suggest that PL is a potential natural anticancer agent for cervical cancer treatment.

Asparanin A inhibits cell migration and invasion in human endometrial cancer via Ras/ERK/MAPK pathway

Asparanin A (AA), a natural compound present in vegetables and medicinal herbs like Asparagus officinalis L., has been investigated extensively for its pharmacological attributes. So far, the effect of AA on endometrial cancer (EC) cell migration and invasion has not been explored. Herein, we elucidated the anti-metastasis mechanism of AA on Ishikawa cells based on miRNA-seq and mRNA-seq integrated analyses. AA treatment led to altered miRNAs expression in Ishikawa cells and inhibited the cell wound healing, cell migration and invasion. Gene Ontology and KEGG enrichment analyses showed that the target genes of different expression miRNAs were significantly enriched in Ras, Rap1 and MAPK signaling pathways. Further verification of these changes via qRT-PCR and Western blot assays in vitro and in vivo demonstrated that AA could suppress human EC cell migration and invasion through Ras/ERK/MAPK pathway. Furthermore, top two miRNAs (miR-6236-p5 and miR-12136_R+8) and top three target genes (KITLG, PDGFD, and NRAS) were identified as functional hub miRNAs and genes through miRNA-target gene network analysis. Our data presented a holistic approach to comprehend the anti-metastatic role of AA in EC after in vitro and in vivo analyses.

Plumbagin induces Ishikawa cell cycle arrest, autophagy, and apoptosis via the PI3K/Akt signaling pathway in endometrial cancer

Plumbagin (PLB) is a naphthoquinone endowed with potential medicinal properties, including anticancer activities. We evaluated the effects of PLB on the viability, cell cycle, autophagy, and apoptosis of endometrial carcinoma Ishikawa cells. The proliferation of cells was significantly inhibited by PLB at 0, 8, 10, and 12 μM. By up regulating the expression of p53 and p21, PLB could block the cell cycle in G2/M phase and down regulate cyclin dependent kinase. The apoptosis in the cancer cells was characterized by noticeable chromatin edge collection, nuclear membrane expansion, and vacuolization. PLB could significantly induce autophagy in cells, and its inhibition ability and apoptosis induction were weakened by the autophagy inhibitor SBI-0206965. Our study suggested that PLB may exert anticancer effects by abrogating PI3K/Akt pathway, which recommends it as a promising future phytotherapeutic candidate for EC treatment.