Toxicology Research最新文献

{"title":"Exploring the Ca²⁺ signaling and cytotoxicity induced by the alantolactone in breast cancer cells and its potential implications in treatment using the Ca²⁺ chelating agent BAPTA-AM.","authors":"Chun-Lang Su, Po-Min Chang, Wei-Zhe Liang","doi":"10.1093/toxres/tfaf044","DOIUrl":"10.1093/toxres/tfaf044","url":null,"abstract":"<p><p>Alantolactone, a bioactive sesquiterpene lactone derived from the roots of <i>Inula helenium</i> (elecampane), has garnered attention in biomedical and pharmacological research for its diverse therapeutic properties, including anticancer, anti-inflammatory, antimicrobial, and antioxidant activities. Despite its well-documented bioactivity, the effects of alantolactone on calcium ion (Ca²⁺) signaling and the underlying mechanisms in human breast cancer cells remain poorly understood. This study explored how alantolactone influences intracellular Ca²⁺ levels ([Ca²⁺]_i), cell viability, and the role of Ca²⁺-dependent pathways in T-47D human breast cancer cells. Specifically, it examined the relationship between Ca²⁺ signaling and cytotoxicity in cells exposed to alantolactone, with or without the Ca²⁺ chelator BAPTA-AM. The findings reveal that alantolactone (25-75 μM) increases [Ca²⁺]_i in a concentration-dependent manner, while concentrations of 25-100 μM induce cytotoxicity, an effect that can be reversed by BAPTA-AM pre-treatment. Removing extracellular Ca²⁺ significantly inhibits Ca²⁺ influx, and both SKF96365 and 2-APB, modulators of store-operated Ca²⁺ channels, block the alantolactone-induced Ca²⁺ entry. Additionally, in a Ca²⁺-free environment, thapsigargin, an inhibitor of the endoplasmic reticulum Ca²⁺ pump, suppresses the alantolactone-induced rise in [Ca²⁺]_i, while alantolactone reduces the [Ca²⁺]_i increase triggered by thapsigargin. Moreover, inhibiting phospholipase C (PLC) with U73122 abolishes the alantolactone-induced [Ca²⁺]_i elevation. These results suggest that alantolactone-induced cell death in T-47D cells is Ca²⁺-dependent, involving Ca²⁺ entry via store-operated channels and Ca²⁺ release from the endoplasmic reticulum, with PLC playing a pivotal role. Importantly, the ability of BAPTA-AM to reverse alantolactone's cytotoxic effects highlights its potential therapeutic significance in breast cancer research.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf044"},"PeriodicalIF":2.2,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12061657/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143957535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"L-asparaginase from the mangrove endophyte <i>Penicillium citrinum</i> MEF 455: a focus on cancer surveillance gene expression in tumor cell lines HL-60 and NCI-H 460.","authors":"Neema Job, K S Sruthy, Divya Jose, Jayesh Puthumana, Manomi Sarasan, K G Nevin, I S Bright Singh, Rosamma Philip","doi":"10.1093/toxres/tfaf067","DOIUrl":"10.1093/toxres/tfaf067","url":null,"abstract":"<p><p>Marine endophytic fungi serve as a valuable source of bioactive molecules, with growing applications in enzyme production. This study investigates the therapeutic potential of glutaminase- and urease-free Type II L-asparaginase derived from the mangrove endophyte <i>Penicillium citrinum</i> MEF 455 against neoplastic cells. Extracellular L-asparaginase production was done using Czapek Dox broth amended with L-asparagine and a 66 kDa molecular mass asparaginase could be observed. The specific activity of 41.6 U/mg with 5.8-fold purification was attained using DEAE cellulose and Sephacryl S-200 column. The kinetic study showed that the <i>Km</i>, <i>Vmax_,</i> and <i>Kcat</i> were 1.370 mM, 161.29 U/mL/min, and 1240.69/min, respectively. Purified L-asparaginase displayed optimal activity at 40 °C and pH 8, with a substrate concentration of 2.5 mM L-asparagine. Metallic ions like Na⁺, K⁺, Mg²⁺, Co²⁺, and Li⁺, improved asparaginase activity. The enzyme displayed strong anticancer potential with considerable reduction in the growth of HL-60, and NCI-H 460 cells with IC₅₀ values of 0.37 ± 0.225 U/mL and 0.39 ± 0.176 U/mL, respectively. Major cancer-controlling genes i.e. p53, caspase-3, caspase-9, NF-kB, Bax, and Rb1 were up-regulated. In contrast, anti-apoptotic i.e. Cox-2 and Bcl-2 were down-regulated on asparaginase treatment in Human cancer cell lines HL-60 and NCI-H 460. The experimental study demonstrates that Type II L-asparaginase produced from an endophytic fungal source, <i>P. citrinum</i> MEF 455, was free from glutaminase and urease activity, thereby minimizing associated immunogenic complications. In general, understanding the physicochemical properties and functionality of the enzyme highlights its potential as a promising antitumor candidate for therapeutic development and clinical applications.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf067"},"PeriodicalIF":2.2,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12089984/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144126126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Role of endoplasmic reticulum stress on formaldehyde-exacerbated allergic asthma in mice.","authors":"Yuchao Zhang, Yutong Guo, Liangyu Liu, Wei Xia, Yi Wang, Mengyin Liu, Dayuan Lin, Jiaxing Li, Yuanchao Zhao, Sijie Zhu, Xudong Liu","doi":"10.1093/toxres/tfaf066","DOIUrl":"10.1093/toxres/tfaf066","url":null,"abstract":"<p><p>Both epidemiological and laboratory evidence indicate a significant relationship between formaldehyde (FA) exposure and allergic asthma. However, the mechanisms underlying the relationship remain unclear. Research has demonstrated that endoplasmic reticulum (ER) stress is closely associated with the onset of allergic asthma. Nonetheless, it has yet to be established whether FA exposure exacerbates allergic asthma by activating ER stress. To systematically investigate the exacerbation of allergic asthma-like symptoms due to FA exposure (0.5 mg/m³) in Balb/c mice, we assessed lung function and histopathology, serum immunoglobulin levels, neuropeptide substance P (SP) and calcitonin gene-related peptide (CGRP) levels, Th2 (IL-4, IL-5, IL-13) and Th17 (IL-22, IL-17A) cytokine levels and biomarkers of the ER stress pathway (IRE1α, PERK, and ATF-6). Additionally, we employed the ER stress antagonist phenylbutyric acid (4-PBA) to confirm the mediating role of ER stress in FA-aggravated allergic asthma. Our findings suggest that prolonged exposure to FA increases levels of ER stress markers, SP, CGRP, Th2 and Th17 cytokines, and immunoglobulin, leading to increased airway mucus hyperplasia and airway remodeling. Furthermore, we demonstrated that blocking ER stress with 4-PBA effectively alleviated associated allergic asthma-like symptoms. In conclusion, we provide evidence that the ER stress signaling pathway plays a significant role in the exacerbation of allergic asthma due to FA exposure.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf066"},"PeriodicalIF":2.2,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12057736/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143955405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Abnormal chenodexycholic acid metabolism programming promotes cartilage matrix degradation in male adult offspring rats induced by prenatal caffeine exposure.","authors":"Bin Li, Hui Gao, Hao Xiao, Hangyuan He, Qubo Ni, Qingxian Li, Hui Wang, Liaobin Chen","doi":"10.1093/toxres/tfaf063","DOIUrl":"10.1093/toxres/tfaf063","url":null,"abstract":"<p><p>Epidemiological evidence links osteoarthritis to fetal origins. Our study shows prenatal caffeine exposure (PCE) in rats predisposes adult offspring to osteoarthritis, associated with elevated intrauterine glucocorticoid levels. Previous research indicates that chenodeoxycholic acid (CDCA), a bile acid, can slow osteoarthritis progression when administered intra-articularly. This study explored if disrupted bile acid metabolism in cartilage affects osteoarthritis risk in adult offspring with PCE. Our findings indicate that the expression of MMP3/MMP13 was upregulated, while endogenous CDCA levels were reduced in the cartilage of PCE-exposed offspring. Furthermore, we observed a persistent reduction in H3K27ac levels at the CYP7B1 promoter and its expression in the cartilage of PCE offspring from fetus to adulthood. Moreover, a sub-physiological level of CDCA promoted NF-κB phosphorylation and the expression of MMP3/MMP13 in chondrocytes <i>in vitro</i>. High levels of glucocorticoids reduced H3K27ac levels and CYP7B1 expression in the promoter region of CYP7B1 through the glucocorticoid receptor and histone deacetylase 4, consequently leading to decreased CDCA levels. In summary, our findings suggest that intrauterine low-expression programming of CYP7B1, induced by elevated glucocorticoid levels, reduces local CDCA levels in the cartilage of PCE offspring, ultimately leading to increased matrix degradation and susceptibility to osteoarthritis.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf063"},"PeriodicalIF":2.2,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12051868/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143953241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cigarette butts: a source of toxicity and potential disruption of <i>Pila virens</i> antioxidant mechanisms.","authors":"Koigoora Srikanth, Sadhanala Siksha Sravani, Rajesh Pamanji, Nadiya Shaik, Gisha Sivan, Nadakuditi Venkata Raju","doi":"10.1093/toxres/tfaf056","DOIUrl":"10.1093/toxres/tfaf056","url":null,"abstract":"<p><p>Cigarette butts (CBs) have emerged as one of the most significant pollutants affecting the environment, particularly aquatic ecosystems. With trillions of cigarettes being manufactured worldwide each year, the disposal of discarded cigarette butts has become a major environmental issue. These butts contain various toxic substances such as nicotine, heavy metals, and other chemicals that can leach into water bodies, posing serious risks to aquatic organisms and disrupting the delicate balance of aquatic life. In this context, freshwater snails, specifically <i>Pila virens</i>, were selected as a model organism to evaluate the impact of CBs on aquatic health. The study exposed <i>P. virens</i> to varying concentrations of CBs-10, 25, and 50%-for different exposure durations (24 h and 48 h). The investigation revealed significant changes in key biomarkers related to oxidative stress and neurotoxicity, including a reduction in glutathione sulfotransferase (GST), protein carbonyl (PC), reduced glutathione (GSH), lipid peroxidation (LPO), glutathione peroxidase (GPx) and Acetylcholine esterase (AChE), respectively. These alterations suggest that CBs induce oxidative stress and neurotoxicity in <i>P. virens</i>, impairing their cellular defense mechanisms. The findings highlight the harmful ecological effects of CBs pollution, emphasizing the urgent need to address this growing environmental concern and its potential consequences on freshwater life. The study contributes valuable insights into the ecotoxicology of CBs contamination in aquatic environments.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf056"},"PeriodicalIF":2.2,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12051869/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143956174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancements in toxicological risk assessment: integrating Ferguson's principle, computational models, and drug safety guidelines, a comprehensive framework for improving risk assessment and resource management in toxicology.","authors":"Saurabh Dilip Bhandare","doi":"10.1093/toxres/tfaf065","DOIUrl":"10.1093/toxres/tfaf065","url":null,"abstract":"<p><p>This investigative study examines the transfer of maternal medications into breast milk and their potential impact on breastfeeding infants. Significant factors influencing drug transfer, including physiochemical properties and milk composition, are analysed to corroborate judicious drug administration in nursing mothers. The study investigates, evaluates, and interprets drugs such as: H|chlorpromazine (New England Nuclear [NEN]), diazepam Roche, C|diclofenac (Ciba-Geigy, 6.6 mCi/mmol, K-277), diclofenac (Ciba-Geigy, 0.1317), digoxin (Wellcome, 11725), fluphenazine (Squibb 12240), phenytoin (NEN, 46 Ci/mmol, 2315-061), phenytoin (Parke-Davis 5419972), pirenzepine (Boehringer-Ingelheim-660206), H|prednisolone (Amersham, 67.4 Ci/mmol, 88), warfarin (Amersham, 46 mCi/mmol, 30), outlining and assessing their transferability and perils notably presented. Ferguson's principle was leveraged to predict drug toxicity, specifically for central nervous system depressants, elucidating drug lethality and safety evaluation. On top of that, advancements in toxicological risk assessment were evaluated, articulated as focusing on naloxone programs, predictive modelling, quantitative structure-activity relationship (QSAR) applications, toxicogenomics, and ordinary differential equation (ODE) models. The comparison between risk assessments and biological monitoring highlights the prominence of evaluating internal dosages. Progress in 3D-QSAR modelling augmented its role in forecasting chemical toxicity, while advancements in toxicogenomics and the application of ODE models have contributed to toxicological research. Hence, the shift toward alternate toxicity assessment methodologies was driven by ethical concerns, budgetary limits, and the demand for more human-relevant data without sacrificing an animal life, which was a concern of the present scientific investigation; fixed by machine algorithms, e.g. random forest, Support Vector Machine (SVM), Ferguson's principle, etc.; an omics data set for correlation through tactile programmed computational heuristics for decision science.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf065"},"PeriodicalIF":2.2,"publicationDate":"2025-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12050033/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143957130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Radiation-induced esophagitis and lung injury during esophageal squamous cell cancer therapy is correlated to tumor gene expression phenotype.","authors":"Wenwen Xu, Yi Wang, Congshu Zhang, Yuqing Chai, Junfeng Gao, Zheng Cao, Yunhong Xia, Hongxia Li","doi":"10.1093/toxres/tfaf062","DOIUrl":"10.1093/toxres/tfaf062","url":null,"abstract":"<p><p>Radiation esophagitis (RE) and Radiation-induced lung injury (RILI) are the main side effects of radiotherapy for esophageal squamous cell cancer (ESCC), which seriously affect the quality of life and therapeutic effect of patients. Then, how to reduce the incidence of RE and RILI is an important topic. We try to establish RE and RILI's prediction scheme based on the gene expression patterns in tumor tissues from patients with ESCC. A total of 37 patients who pathological preliminary diagnosed as ESCC and received radical radiotherapy from 2016 January 1 to 2019 December 31 were enrolled in this study. Use 3-plex qPCR to detect gene expression in ESCC. Our results showed that gene expressions in the Mitogen-activated protein (MAP) kinase signaling (HRAS, MAP2K1, MAPK1, CRAF and KRAS) were positively related to Severe RE (SRE), while Fibroblast growth factor (FGF) signaling showed a negative correlation. We established a c-Index calculation model to predict SRE. Receiver operating characteristic curve were applied to determine the prognostic value of the risk model. Besides, patients with SRE seem to be more easily to develop higher-level of RILI. Taken together, we constructed a novel radiotherapy response-related gene signature, which may be developed into a powerful tool for forecasting the risk of SRE in ESCC radiotherapy patients.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf062"},"PeriodicalIF":2.2,"publicationDate":"2025-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12050034/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143953656","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effect of 50 Hz magnetic fields on cellular sensitivity of mouse spermatogenic cell lines to hydrogen peroxide.","authors":"Xiaoxia Wei, Longtao Zhu, Ying Zhu, Xinyuan Zhao, Chuan Sun, Guangdi Chen","doi":"10.1093/toxres/tfaf059","DOIUrl":"10.1093/toxres/tfaf059","url":null,"abstract":"<p><p>With the widespread application of electromagnetic technology, electromagnetic fields (EMFs) emitted from various electric and electronic devices have significantly altered the electromagnetic environment. This has raised concerns about the potential health impacts of EMFs. Previous studies have indicated that EMFs may influence male infertility, with oxidative stress proposed as a key factor; however, the underlying mechanisms remain unclear. In this study, we aimed to determine whether EMFs enhance the impact of oxidative stress on male infertility. We investigated the effects of 50 Hz magnetic fields (MFs) on the sensitivity of mouse spermatogenic cell lines (GC-1 spg and GC-2 spd) to low-dose hydrogen peroxide (H₂O₂, 5 and 10 μM). Our findings revealed that pre-exposure to 2.0 mT 50 Hz MFs for 24 h increased the sensitivity of GC-2 spd cells to low-dose H₂O₂ in terms of γH2AX foci formation, a marker for DNA damage repair. However, no significant changes were observed in DNA fragmentation, cell viability, or cell cycle progression in either GC-1 spg or GC-2 spd cells. In conclusion, our results suggest that 50 Hz MFs do not significantly enhance the sensitivity of mouse spermatogenic cell lines to low-dose H₂O₂.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 3","pages":"tfaf059"},"PeriodicalIF":2.2,"publicationDate":"2025-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12049193/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143957370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chaperone mediated autophagy modulates microglia polarization and inflammation via LAMP2A in ischemia induced spinal cord injury.","authors":"Dan Fu, Ziyou Li, Huafeng Feng, Fangling Fan, Wang Zhang, Liang He","doi":"10.1093/toxres/tfaf061","DOIUrl":"10.1093/toxres/tfaf061","url":null,"abstract":"<p><p>Spinal cord injury (SCI)-induced ischemic delayed paralysis is one of the most serious side effects of aneurysms surgeries. Recent studies prove that the activation of autophagy, including macroautophagy and micro-autophagy pathways, occur during SCI-induced brain neuron damage. However, the role of chaperone mediated autophagy (CMA) during SCI remains to be unveiled. In the present work, rat model of delayed paralysis after aneurysms operation and adenovrius induced LAMP2A knockdown in microglia cells were applied in the present work to investigate the involvement of LAMP2A-mediated CMA in the aneurysm operation related SCI and delayed paralysis. The results showed that LAMP2A was upregulated in the SCI procedure, and contributed to neuron death and pro-inflammation perturbation via inducing iNOS⁺ polarization in microgila. We additionally observed that knockdown of LAMP2A resulted in the shift of microglia from iNOS⁺ to ARG1⁺ phenotype, as well as alleviated neuron damage during SCI. Furthermore, the analysis of BBB score, the result of immunohistological staining, and protein detection confirmed the activation of LAMP2A-mediated CMA activation and its interaction with NF-κB signaling, which leads to neuron death and motor function loss. These results prove that LAMP2A-mediated CMA contributes to the upregulation of pro-inflammatory cytokines and results in cell death in neurons during ischemic delayed paralysis via activating NF-κB signaling. Inhibition of LAMP2A promotes neurons survival during ischemic delayed paralysis.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 2","pages":"tfaf061"},"PeriodicalIF":2.2,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12038812/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143957392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FTO-mediated m6A demethylation of KLF4 promotes the proliferation and collagen deposition of keloid fibroblasts.","authors":"Yanqi Li, Wanchao Wang, Yuge Wang, Hongmei Ai","doi":"10.1093/toxres/tfaf058","DOIUrl":"10.1093/toxres/tfaf058","url":null,"abstract":"<p><p>This study aims to elucidate the molecular mechanism mechanism by which FTO affects fibroblast proliferation and collagen deposition in keloids. Human keloid fibroblasts (KFs) and normal fibroblasts were cultured in vitro. FTO expression was silenced in KFs, and cell viability and proliferation were evaluated via CCK-8 and clone formation assays. FTO, KLF4, and MC1R expressions were quantified via qRT-PCR, while the protein levels of FTO, KLF4, MC1R, Collagen I, and Collagen III were determined by Western blot. The m⁶A RNA methylation status of total RNA was evaluated using the EpiQuik m6A RNA Methylation Quantification Kit. Post-actinomycin D treatment, the stability of KLF4 mRNA and its m⁶A modification level were measured. ChIP and dual-luciferase reporter assays confirmed the binding between KLF4 and MC1R promoter. KFs presented with significantly enhanced proliferation and collagen deposition, correlating with elevated FTO expression. Silence of FTO repressed the proliferation and collagen deposition of KFs, and elevated the m6A levels of total RNA and KLF4 mRNA in KFs, resulting in enhanced KLF4 mRNA stability and expression. KLF4 bound to the MC1R promoter and promoted MC1R expression. In conclusion, FTO represses KLF4 expression by removing m6A modification and further diminishes MC1R expression, thereby facilitating KF proliferation and collagen deposition.</p>","PeriodicalId":105,"journal":{"name":"Toxicology Research","volume":"14 2","pages":"tfaf058"},"PeriodicalIF":2.2,"publicationDate":"2025-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12031721/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143957069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}