4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a tobacco-specific carcinogen, closely linked to lung cancer development. Cancer stem cells (CSCs) contribute to malignant tumorigenesis and progression. Long nonco...4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a tobacco-specific carcinogen, closely linked to lung cancer development. Cancer stem cells (CSCs) contribute to malignant tumorigenesis and progression. Long noncoding RNAs (lncRNAs) play a crucial role in regulating cancer stemness, yet the roles and mechanisms remain unclear. In our previous study, we established a malignantly transformed cell line (Beas-2B-NNK) by chronic administration of low-dose (5 μM) NNK to human normal lung epithelial cells (Beas-2B). We identified a candidate lncRNA WFDC21P through high-throughput sequencing and bioinformatics analysis. Cancer-like properties were evaluated through CCK-8 assay, wound healing, soft agar colony formation, and nude mouse xenograft experiments. Cancer stemness was determined via the suspension sphere assay, the mRNA and protein levels of stemness markers were detected using qRT-PCR and Western blot. Knockdown and overexpression experiments confirmed the function of WFDC21P and its downstream regulatory pathways. Our results indicate that Beas-2B-NNK cells show enhanced cancer-like phenotypes and high expression of lncRNA WFDC21P compared with Beas-2B cells. Knockdown of WFDC21P inhibits proliferation, migration, and tumor stemness, while overexpression of WFDC21P promotes these phenotypes. Mechanistically, WFDC21P mediates these effects by targeting the TGF-β signaling pathway. In conclusion, WFDC21P regulates tumor stemness via the TGF-β pathway to mediate NNK-induced lung epithelial cell malignant transformation.
Dibutyl phthalate (DBP) has been recognized as an endocrine disrupting chemical, which poses significant threats to male reproductive health, though its precise mechanism of testicular toxicity has not been fully clarifi...Dibutyl phthalate (DBP) has been recognized as an endocrine disrupting chemical, which poses significant threats to male reproductive health, though its precise mechanism of testicular toxicity has not been fully clarified. We combined in vivo and in vitro experiments with transcriptome sequencing to investigate the toxicity of DBP and its active metabolite monobutyl phthalate (MBP) in prepubertal male mice and Sertoli cells, and explored the underlying molecular mechanisms. The results showed that DBP affected testicular development and disrupted the blood-testis barrier (BTB) in mice. Through bioinformatic analysis, we validated that MBP promoted G1 phase cell cycle arrest, apoptosis, and DNA damage in Sertoli cells. Furthermore, the increased expression of p-ATM, p-Chk2, p-p53 indicated that the ATM/Chk2/p53 signaling pathway was activated upon MBP treatment. Inhibition of p53 or ATM expression partially reversed the pro-apoptotic and cell cycle-arrested phenotypes, whereas p53 overexpression reverted these phenotypic abnormalities. These findings indicated that prepubertal DBP exposure impaird BTB integrity and induced DNA damage in association with pathological activation of the ATM/Chk2/p53 signaling pathway, thereby contributing to Sertoli cell apoptosis and proliferative arrest.
T-2 toxin (T-2) and HT-2 toxin (HT-2) are trichothecene mycotoxins frequently detected in cereals. While exposure has primarily been evaluated using dietary assessment methods, human biomonitoring provides an alternative...T-2 toxin (T-2) and HT-2 toxin (HT-2) are trichothecene mycotoxins frequently detected in cereals. While exposure has primarily been evaluated using dietary assessment methods, human biomonitoring provides an alternative approach to assess internal exposure. This follow-up research aimed to further characterise the toxicokinetics of the mycotoxins T-2 and HT-2 and to subsequently develop a physiologically based kinetic (PBK) model, enabling prediction of urinary biomarker concentrations based on external exposure and vice versa. To this end, urine samples from 40 Norwegian adults were analysed for T-2 and its metabolites using targeted and suspect screening analytical approaches. Dietary exposure of these participants was estimated using 24-h weighed food records combined with Norwegian occurrence data. In parallel, toxicokinetic information on the mycotoxins was generated using in vitro liver microsome experiments and a literature review. Total HT-2 (HT-2 and its glucuronides) was detected in 97% of the 24-h urine voids, whereas total T-2 and T-2 triol were not detected. Suspect screening tentatively identified eight mono- and di-hydroxylated metabolites of T-2 and HT-2. In vitro, 98% of T-2 was metabolised within 2 h, with HT-2 accounting for 97% of the metabolites. Following a 2 h incubation with HT-2, 73% of the added HT-2 remained unchanged, and a large fraction was unidentified compounds (26%). Parametrisation of the PBK model was supported by data from public literature. This study advances the understanding of the human toxicokinetics of T-2 and HT-2, and demonstrates the utility of PBK modelling to link external and internal exposure. The identification of multiple hydroxylated urinary metabolites suggests that HT-2 alone does not fully represent internal exposure. This research provides direction for future metabolism studies which are needed to support further development of a robust PBK model for T-2 and HT-2.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Exposure to persistent organic pollutants such as polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) and polychlorinated biphenyls (PCBs) could promote several adverse health effects. This study aimed to quantify the...Exposure to persistent organic pollutants such as polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) and polychlorinated biphenyls (PCBs) could promote several adverse health effects. This study aimed to quantify the relative contributions of various exposure pathways to the body burden of PCDD/Fs and PCBs. We used a physiologically based pharmacokinetic model to simulate the internal exposure dynamics across different ages and for different exposure scenarios. We incorporated various parameters into the model to account for exposure through ingestion. These included dietary factors (vegetarian or mixed diet and consumption of home-produced eggs from contaminated areas), breastfeeding duration, maternal age, and soil concentrations. The modelled results from the simulated serum concentration showed that dietary intake, particularly from animal-fat-rich foods produced on contaminated areas, constitutes the predominant source of exposure. Breastfeeding significantly impacts early-life body burden temporarily, while soil and dust ingestion contribute minimally, even in the presence of high PCDD/F concentrations in soil. When an average diet is consumed in conjunction with six months of breastfeeding, the No-Observed-Adverse-Effect Level is not reached at 9 years old for PCDD/Fs WHO-TEQ-05 concentration. These findings highlight the need for targeted public health recommendations, such as reducing animal fat consumption, to mitigate the population's overall body burden.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Phthalates (PAEs) are widely used plasticizers that are increasingly linked to tumorigenesis. In this study, we integrated network toxicology and machine learning to elucidate the potential mechanisms of PAEs in lung squ...Phthalates (PAEs) are widely used plasticizers that are increasingly linked to tumorigenesis. In this study, we integrated network toxicology and machine learning to elucidate the potential mechanisms of PAEs in lung squamous cell carcinoma (LUSC). By integrating multi-database target prediction, The Cancer Genome Atlas Program (TCGA) transcriptomic analysis, and algorithmic screening, four key genes, ANXA5, CDK4, MMP1, and SRD5A1, were identified as major targets of PAEs. Molecular docking and dynamics simulations indicated stable binding of butyl benzyl phthalate (BBP) to ANXA5, and SRD5A1. In vitro validation using H226 and NCI-H1703 cells confirmed these predictions. BBP at non-cytotoxic concentrations (0.1-10 μM) altered cell behavior: 0.1 μM BBP promoted migration and upregulated SRD5A1 and ANXA5, whereas 10 μM BBP downregulated ANXA5, showing a non-monotonic "low-dose activation-high-dose inhibition" pattern typical of endocrine disruptors. Collectively, these results indicate that BBP interferes with LUSC progression through multiple target and multipathway mechanisms, including direct binding to key proteins, disturbance of steroid metabolism, and modulation of membrane signaling. This integrative study links environmental phthalate exposure with lung cancer pathogenesis and provides a hypothesis worthy of further investigation for risk assessment and targeted prevention.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Ethyl palmitate was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, photoirritation/photoallergenicity, skin sensitization, and environmental safety. Data from read-...Ethyl palmitate was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, photoirritation/photoallergenicity, skin sensitization, and environmental safety. Data from read-across analogs methyl hexadecanoate (CAS # 112-39-0) and methyl decanoate (CAS # 110-42-9) show that ethyl palmitate is not expected to be genotoxic. Data on read-across analog ethyl octadecenoate (CAS # 111-61-5) provide a calculated Margin of Exposure (MOE) > 100 for the repeated dose toxicity and reproductive toxicity endpoints. Data from read-across analog methyl hexadecanoate (CAS # 112-39-0) provided ethyl palmitate a No Expected Sensitization Induction Level (NESIL) of 2400 μg/cm for the skin sensitization endpoint. The photoirritation/photoallergenicity endpoints were evaluated based on ultraviolet/visible (UV/Vis) spectra; ethyl palmitate is not expected to be photoirritating/photoallergenic. The local respiratory toxicity endpoint was evaluated using the Threshold of Toxicological Concern (TTC) for a Cramer Class I material, and the exposure to ethyl palmitate is below the TTC (1.4 mg/day). The environmental endpoints were evaluated; ethyl palmitate was found not to be Persistent, Bioaccumulative, and Toxic (PBT) as per the International Fragrance Association (IFRA) Environmental Standards, and its risk quotients (RQs), based on its current volume of use (VoU) in Europe (EU), North America (NA), Asia-Pacific (AP), and South America (SA) (i.e., Predicted Environmental Concentration/Predicted No Effect Concentration [PEC/PNEC]), are <1.
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Api AM, Bartlett A, Belsito D
… +31 more, Botelho D, Bruze M, Bryant-Friedrich A, Burton GA, Cancellieri MA, Chon H, Cronin M, Crotty S, Dagli ML, Dekant W, Deodhar C, Farrell K, Fryer AD, Jones L, Joshi K, Lapczynski A, Laskin DL, Lavelle M, Lee I, Moustakas H, Muldoon J, Penning TM, Piersma AH, Ritacco G, Sadekar N, Schember I, Schultz TW, Siddiqi F, Sipes IG, Sullivan G, Thakkar Y
Particulate matter 2.5 (PM2.5) is a major environmental pollutant linked to neurological disorders through poorly understood mechanisms. We investigated whether PM2.5 induces disulfidptosis, a novel form of regulated cel...Particulate matter 2.5 (PM2.5) is a major environmental pollutant linked to neurological disorders through poorly understood mechanisms. We investigated whether PM2.5 induces disulfidptosis, a novel form of regulated cell death characterized by NADPH depletion and disulfide stress, in neuronal cells. Combining bioinformatics analysis of a PM2.5-exposed human cohort (GSE60767, n = 466) with in vitro experiments in primary neurons and in vivo validation in rats, we identified a characteristic molecular signature featuring SLC3A2 upregulation and OXSM downregulation (p < 0.01). This imbalance was associated with significant immune dysregulation (elevated memory B cells, CD8 T cells, and Tregs; reduced monocytes) and metabolic disturbances (NADP+/NADPH imbalance, cystine overload). Direct evidence from non-reducing Western blot confirmed disulfide-mediated actin crosslinking in both cultured neurons and hippocampal tissues of PM2.5-exposed rats. Lentiviral manipulation revealed a unidirectional regulatory relationship wherein SLC3A2 knockdown (RNAi-SLC3A2) increased OXSM expression, while OXSM overexpression (OE-OXSM) did not affect SLC3A2 levels. Both RNAi-SLC3A2 and OE-OXSM interventions improved neuronal survival and normalized NADP+/NADPH ratios after PM2.5 exposure (200 μg/mL). However, RNAi-SLC3A2 significantly inhibited cystine uptake. Correspondingly, RNAi-SLC3A2 demonstrated more robust neuroprotection compared to OE-OXSM, as evidenced by greater improvements in neuronal viability and complete restoration of action potential amplitude. Collectively, these findings establish that PM2.5 induces neuronal disulfidptosis through hierarchical SLC3A2-OXSM axis dysregulation and identify this unidirectional relationship as a novel therapeutic target for PM2.5-associated neurological injury.