ARCHIVES OF TOXICOLOGY
·
2022
·Vol. 97
·期 2
·441-456
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IF 6.168
Xu Chi,Jiang Shuyun,Ma Xingyu,Jiang Zeyong,Pan Youguang,Li Xiang,Zhang Liying,Zhou Hao,Chen Shen,Xing Xiumei,Chen Liping,Fu Wenfan,Wang Qing,Chen Wen,Li Daochuan
Cisplatin is recommended as a first-line chemotherapeutic agent against advanced non-small cell lung cancer (NSCLC), but acquired resistance substantially limits its clinical efficacy. Recently, DNA methylation has been identified as an essential contributor to chemoresistance. However, the precise DNA methylation regulatory mechanism of cisplatin resistance remains unclear. Here, we found that nicotinamide nucleotide transhydrogenase ( NNT ) was silenced by DNA hypermethylation in cisplatin resistance A549 (A549/DDP) cells. Also, the DNA hypermethylation of NNT was positively correlated to poor prognosis in NSCLC patients. Overexpression of NNT in A549/DDP cells could reduce their cisplatin resistance, and also suppressed their tumor malignancy such as cell proliferation and clone formation. However, NNT enhanced sensitivity of A549/DDP cells to cisplatin had little to do with its function in mediating NADPH and ROS level, but was mainly because NNT could inhibit protective autophagy in A549/DDP cells. Further investigation revealed that NNT could decrease NAD + level, thereby inactivate SIRT1 and block the autophagy pathway, while re-activation of SIRT1 through NAD + precursor supplementation could antagonize this effect. In addition, targeted demethylation of NNT CpG island via CRISPR/dCas9-Tet1 system significantly reduced its DNA methylation level and inhibited the autophagy and cisplatin resistance in A549/DDP cells. Thus, our study found a novel chemoresistance target gene NNT , which played important roles in cisplatin resistance of lung cancer cells. Our findings also suggested that CRISPR-based DNA methylation editing of NNT could be a potential therapeutics method in cisplatin resistance of lung cancer.
研究领域:
肿瘤学
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分子生物学
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表观遗传学
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产品关联
DOI
PubMed
高影响因子
Animal Nutrition
·
2025
·
IF 7.5
Xinyao Zhang,Ke Cheng,Lixue Dong,Dexing Zhu,Qingyong Li,Zhongbao Guo,Yongju Luo,Apeng Liu,Juan Tian,Di Peng,Jianmin Zhang,Yangyang Liu,Hua Wen,Ming Jiang
Fava bean is currently utilized as a specialized feed ingredient to enhance the muscle texture of fish. When incorporated at appropriate levels, fava beans can improve fish muscle crispy texture. However, high dietary levels of fava beans may lead to liver damage in fish, although the specific mechanisms behind this effect remain unclear. In this study, 600 genetic improvement of farmed tilapia (GIFT; Oreochromis niloticus ) of uniform size (initial body weight 552.3 ± 10.12 g) and healthy appearance were randomly divided into four groups, each with three replicates of 50 fishes. The fish diets contained different levels of fava beans (0, 50%, 60%, and 70%), designated as D0, D50, D60, and D70, respectively, over a ten-week feeding experiment. A comprehensive analysis of the impact of fava beans on muscle texture and liver health in GIFT was performed. The results showed that 50%, 60%, and 70% dietary fava beans had no significant effect on the growth performance of GIFT ( P > 0.05), but 70% dietary fava beans increased feed conversion ratio (FCR) ( P = 0.022). Consumption of fava beans at levels of 50%, 60%, and 70% in the diet altered GIFT muscle texture ( P < 0.05), and adversely effected serum biochemistry and liver health ( P < 0.05). Non-targeted metabolome analysis of the liver revealed that dietary fava beans may cause irreversible damage to liver cells by disrupting the DNA methylation process ( P < 0.05). This finding enhances the understanding of the mechanisms behind the negative impact of fava bean consumption on liver function. It is crucial to investigate potential strategies to mitigate these adverse effects while maintaining the desired muscle quality when fava beans are used in aquatic feed.
研究领域:
水产养殖学
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分子生物学
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营养学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Baoting Dou,Hui Zhou,Xiguang Han,Po Wang
The dissociation of the walking strand from the track gives rise to decreased efficiency and long reaction time of DNA walkers. In this work, we constructed a DNA walker combining the introduction of a wedge segment with a bimetallic metal–organic framework (MOF) electrocatalyst to solve this problem. The target methylated DNA acted as a single-legged walker, and the immobilization probe assembled on the track contained a wedge segment that was complementary to the target methylated DNA persistently, inhibiting its dissociation from the track. The fuel strand modified with a bimetallic MOF would drive the target strand to conduct branch migration and move processively along the track. The stepwise movement of the target strand resulted in the loading of numerous bimetallic MOF catalysts to reduce H2O2 at the electrode interface, thereby a significantly increased current response would be obtained for the detection of methylated DNA. This DNA walker achieved a detection limit of 200 aM within 20 min and effectively distinguished DNA with different methylation statuses, which would pave a way for rapid and sensitive monitoring of DNA methylation.
研究领域:
生物化学
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分析化学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Science China-Chemistry
·
2024
·
IF 10.4
Xu Xing,Zeng Xi,Lin Xin,Lin Shiyan,Liang Shanshan,Tian Tian,Su Rui,Song Jia,Yang Chaoyong
Single-cell joint analysis of methylome and transcriptome reveals how the methylation regulates the transcriptional activity. However, traditional bench-top protocols for single-cell DNA methylation and RNA transcription co-detection are labor-intensive, cost-ineffective and contaminant-prone. Herein, we establish the DMF-scMT-seq, a highly-efficient and cost-effective method to simultaneously analyze single-cell DNA methylation and transcriptional activity based on digital microfluidics. DMF-scMT-seq automates the workflow of single-cell isolation, cellular hypotonic lysis, nucleic acid separation and methylome/transcriptome library construction in a contactless and addressable way. The system ensures high accuracy ( R >0.85), high gene detection ability (14,697 genes per cell at 4 million sequencing depth), and high CpG coverage (677,198 CpG sites per cell at 1 million sequencing depth). By using DMF-scMT-seq, the relationship of DNA methylation and RNA transcription under different genomic contexts is resolved. We further apply DMF-scMT-seq to study the dynamics of transcription regulation with methylation-inhibiting anti-tumor Decitabine, and identify the methylated promoter/gene body driven genes in response to Decitabine treatment. DMF-scMT-seq facilitates the construction of the correlation of DNA methylation and transcriptional activity at the single-cell level in a flexible, sensitive and accurate way, which is anticipated to be a powerful tool in studying single-cell biological systems.
研究领域:
生物信息学
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表观遗传学
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分子生物学
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DOI
高影响因子
Frontiers in Plant Science
·
2017
·
IF 4.291
Zha Liangping,Liu Shuang,Liu Juan,Jiang Chao,Yu Shulin,Yuan Yuan,Yang Jian,Wang Yaolong,Huang Luqi
The content of active compounds differ in buds and flowers of Lonicera japonica (FLJ) and L. japonica var. chinensis (rFLJ). Chlorogenic acid (CGAs) were major active compounds of L. japonica and regarded as measurements for quality evaluation. However, little is known concerning the formation of active compounds at the molecular level. We quantified the major CGAs in FLJ and rFLJ, and found the concentrations of CGAs were higher in the buds of rFLJ than those of FLJ. Further analysis of CpG methylation of CGAs biosynthesis genes showed differences between FLJ and rFLJ in the 5′-UTR of phenylalanine ammonia-lyase 2 (PAL2). We identified 11 LjbZIP proteins and 24 rLjbZIP proteins with conserved basic leucine zipper domains, subcellular localization, and electrophoretic mobility shift assay showed that the transcription factor LjbZIP8 is a nuclear-localized protein that specifically binds to the G-box element of the LjPAL2 5′-UTR. Additionally, a transactivation assay and LjbZIP8 overexpression in transgenic tobacco indicated that LjbZIP8 could function as a repressor of transcription. Finally, treatment with 5-azacytidine decreased the transcription level of LjPAL2 and CGAs content in FLJ leaves. These results raise the possibility that DNA methylation might influence the recruitment of LjbZIP8, regulating PAL2 expression level and CGAs content in L. japonica.
研究领域:
植物学
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分子生物学
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药用植物研究
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DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Hongding Zhang,Sifei Wu,Zhixiao Song,Linxia Fang,Hai-Bo Wang
As a promising biomarker, the level of methylated DNA usually changes in the early stage of the cancer. Ultrasensitive detection of the changes of methylated DNA offers possibility for early diagnosis of cancer. In this work, a tannic acid-accelerated Fenton chemical reaction amplification was firstly proposed for the construction of ultrasensitive fluorescent assay. Tannic acid was used as reductant to accelerate Fenton reaction procedure through the conversion of Fe 3+ /Fe 2+ , generating hydroxyl radicals (·OH) continuously. The produced ·OH oxidized massive non-fluorescent terephthalic acid (TA) to fluorescent-emitting hydroxy terephthalic acid (TAOH). In this way, the fluorescent signal could be greatly enhanced and the sensitivity was improved almost 116 times. The proposed signal amplification strategy was further applied to detect of DNA methylation with the assistance of liposome encapsulated with tannic-Fe 3+ complexes. The methylated DNA was firstly captured through the hybridization with its complementary DNA that were pre-modified in the 96-well plate via the combination between streptavidin (SA) and biotin . Then, 5 mC antibody on the surface of liposomes specially recognized and combined with methylation sites, which brought large amount of tannic-Fe 3+ complexes to participate Fenton reaction. The fluorescence of generated TAOH was depended on the concentration of methylated DNA. The assay showed good analytical performance for methylated DNA with a limit of detection (LOD) of 1.4 fM. It's believed that tannic acid-accelerated Fenton chemical reaction amplification strategy provides a promising platform for ultrasensitive fluorescent detection of low abundant biomarkers.
研究领域:
生物化学
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医学诊断
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分析化学
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产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL PHARMACOLOGY
·
2025
·
IF 5.6
Dan Hu,Xiaoting Wu,Manman Hou,Pin Song,Qu Chen,Yingdong Ni
Ursodeoxycholic acid (UDCA) has been widely used for treatment of biliary diseases with high incidence among pregnant women. However, the effect of maternal UDCA exposure on offspring metabolism particularly on hepatic lipid metabolism has not been reported. Seventeen pregnant Sprague-Dawley rats were administered with low-dosage UDCA (L-UDCA, 250 mg/kg, n = 6), high-dosage UDCA (H-UDCA, 500 mg/kg, n = 6) or phosphate buffered saline (CON, n = 5) during pregnancy. After parturition, pups from each litter were sacrificed for blood and hepatic tissues sampling at 21 days of age. Maternal exposure to H-UDCA increased body weight and liver weight of the offspring before weaning. The levels of triglyceride (TG) in both plasma and liver of offspring were increased in maternal UDCA exposure and led to a greater hepatic lipid deposition in offspring. UDCA treatment predominantly promoted the d e novo lipogenesis (DNL) in offspring liver by upregulating the gene expression of ATP citrate lyase ( Acly ), acetyl-coA carboxylases ( Acc ), and fatty acid synthase ( Fasn ), as well as protein expression of FASN and ACC. The level of fibroblast growth factor 15 (FGF15) in plasma of offspring was increased in UDCA treatment compared to control. Moreover, protein expression of DNA methyltransferase 1 (DNMT1) in the liver of offspring was decreased in a dose-dependent manner and the CpG methylation level in the Fasn promoter region was decreased. This study provides insights into maternal UDCA exposure promoting offspring hepatic lipid deposition, which is associated with hypomethylation modification of cytosine-phosphate-guanine (CpG) islands on the promoter of Fasn gene.
研究领域:
发育毒理学
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表观遗传学
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脂质代谢研究
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围产医学
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药物代谢
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 3
·1-11
·
IF 5.3
Liu Huamin,Li Chenghong,Wang Lina,Fang Lichao,Huang Hui,Deng Jun,Hu Yue,Li Mimi,Ran Xiaoping,Li Lulu,Zheng Junsong
A “signal-off” photoelectrochemical (PEC) sensing platform has been designed for the ultrasensitive detection of DNA methylation levels and multiple methylated sites. The platform employs tungsten trioxide and TpPa-1-COF loaded by gold nanoparticle (AuNPs@WO 3 @TpPa-1-COF) composite material as the photoactive component and p-type reduced graphene (rGO) as an efficient quencher. The PEC signal of AuNPs@WO 3 @TpPa-1-COF composite is effectively quenched in the presence of p-type rGO, because p-type rGO can compete with AuNPs@WO 3 @TpPa-1-COF to deplete light energy and electron donors. In addition, a hybrid strand reaction (HCR) amplification strategy fixes more target DNA and then combines with rGO-modified anti-5-methylcytosine antibody to facilitate ultrasensitive DNA methylation detection. Under optimal conditions, DNA methylation can be measured within a linear concentration range of 10 –14 to 10 –8 M, with an exceptionally low detection limit of 0.19 fM (S/N = 3). At the same time, the platform can conduct quantitative determination of multi-site methylation, with the linear equation △I = 44.19Log A + 61.43, and the maximum number of methylation sites is 5. The sensor demonstrates high sensitivity, excellent selectivity, and satisfactory stability. Furthermore, the proposed signal-off PEC strategy was successfully employed to detect DNA methylation in spiked human serum samples, with recoveries ranging from 93.17 to 107.28% and relative standard deviation (RSD) ranging from 1.15 to 5.49%. Graphical
研究领域:
生物化学
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分析化学
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材料科学
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DOI
PubMed
高影响因子
Cancers
·
2021
·Vol. 13
·期 4
·664
·
IF 6.639
Li-Yue Sun,Zi-Ming Du,Yu-Ying Liu,Yan-Hong Li,Xiao-Min Liu,Ting Wang,Jian-Yong Shao
Simple SummaryPrevious studies have established an electrochemical detection method for the rapid detection of cfDNA (circulating free DNA) methylation and found that this technology might be a potential method for cancer diagnosis. However, the underlying mechanism and its role in the diagnosis and prognosis of malignant tumors are not well-characterized. In present study, we utilized the electrochemical detection method to detect the DNA methylation status by using electron microscopies and infrared spectroscopy and found that DNA with different methylated levels adsorbed to the gold surface differently, which was likely mediated by hydrophobic bonds. In addition, after detection of the cfDNA methylation status from 505 normal individuals, 725 cancer patients before treatment, and 549 patients after treatment, we found that the cfDNA adsorption rate could be used as an indicator for the diagnosis and prognosis prediction of pan-cancer. Our research provides a novel method for the liquid biopsy of cancer for diagnosis and prognosis predictions.AbstractPrior research has established an electrochemical method based on the differential adsorption capacity of gold surfaces with different methylated DNA degrees and found that this method might be valuable for cancer diagnosis by detecting circulating free DNA methylation. However, further investigation on the underlying mechanism and validation of its diagnostic and prognostic values in a large cohort of malignant tumors was limited. We found that DNA with different methylation levels formed particles of diverse sizes on the gold surface. Hydrophobic bonds played a significant role in the binding process of methylated DNA to the gold surface. The detection condition of an adsorption time of 10 min and temperature of 20 °C was optimal. In a large cohort of plasma samples from the patients with different malignant tumors, as well as normal individuals, we found that the electrochemical detection method based on the differential adsorption capacity of methylated DNA degree on a gold surface could be used as a noninvasive tool for malignant tumor diagnosis and prognostic evaluation. The diagnostic efficiency of this method in malignant tumors was even slightly better than that of the current tumor biomarkers widely used in routine clinical practice (circulating free DNA (cfDNA) vs. carcinoembryonic antigen (CEA), 0.8131 vs. 0.7191 and cfDNA vs. CA19-9, 0.7687 vs. 0.6693).Keywords:electrochemical;circulating free DNA methylation;malignant tumor;diagnostic;prognostic
研究领域:
医学
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分子生物学
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肿瘤学
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产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2021
·Vol. 28
·期 23
·30101-30110
·
IF 4.223
Bian Xiaoxue,Gao Yongfei
An unexplored attributing molecular mechanism of Cd toxicity is interference with the epigenetic machinery, such as DNA methylation, processes that are crucial for early fetal development. In order to investigate the effects of Cd on the expression of metallothionein (MT) and Dnmts transcripts, markers of DNA methylation, and signaling pathway gene expression, zebrafish embryos were exposed during 24 hours post-fertilization (starting at maximum 8-cell stage) to 0.0089, 0.089, and 0.89 μM Cd. The results showed that the Cd accumulation in zebrafish embryo reached a stable level after 12 hpf, and the Cd accumulation at individual time points was significantly different among different concentration groups. MT mRNA fold was significantly positive with the Cd content in embryos. We observed that the expression level of DNA methyltransferase ( Dnmts ) in the 0.089 μM Cd exposure group was significantly up-regulated. Dnmt1 expression was significantly up-regulated in the 0.89 μM Cd exposure group, and Dnmt3s expression and global methylation levels were significantly down-regulated. Cd up-regulated ErbB-3 gene expression, down-regulated ErbB-4 gene expression, and neutralized ErbB-1 gene expression. Cd activated Ca 2+ , MAPK-JUK , p38 MAP kinase, PI3K-AKT , and VEGF signaling pathway genes, indicating these pathway genes related to Cd exposure level. The results are helpful to clarify the molecular mechanism of DNA methylation in zebrafish embryo under metal pressure and further interference with the epigenetic machinery.
研究领域:
分子生物学
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环境毒理学
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发育生物学
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产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2025
·
IF 6.1
Danni Jiang,Yang Yang,Xiaojuan Han,Qianni Li,Yingxiu Ma,Jun Jiao,Lan Chao
Fluorene-9-bisphenol (BHPF), a widely recognized alternative to bisphenol A (BPA), has been increasingly used in a wide range of industries. However, despite numerous studies, the direct mechanism underlying its male reproductive toxicity remains poorly understood. Here, a BHPF-exposed mouse model was established to evaluate the effect of BHPF on male fertility. The results showed that BHPF exposure induced reproductive dysfunction, including testis damage, spermatogenesis impairment, steroid hormone disruption and sperm quality degradation. Mechanistically, over 90,000 differentially methylated regions were identified and increased global testicular methylation levels indicated that DNA methylation may be associated with BHPF-induced testicular damage. Transcriptome analysis revealed 221 up-regulated genes and 227 down-regulated genes in BHPF-exposed mice, mainly enriched in the steroid biosynthetic pathway. Further combined methylome and transcriptome analysis revealed the critical methylated genes potentially involved in lipid metabolism. The differentially expressed genes (Cyp4a10 and Rdh1) were further confirmed to correlate with impaired male mouse fertility. Taken together, the results of this study provide a better understanding of the molecular mechanisms of toxicity effects induced by BHPF exposure from the perspective of methylome and transcriptome.
研究领域:
毒理学
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表观遗传学
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生殖生物学
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产品关联
DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2023
·
IF 5
Ying Cheng,Chen Chen,Fang Wang,Zilin Chen
Ten-eleven translocation 1 (TET1) is a member of the TET enzyme family of dioxygenases, which plays an important role in active DNA demethylation. Therefore, the sensitive TET1 detection could help us better understand DNA methylation-demethylation in epigenetics. Here we report a detection method that consists of electrode fabrication, TET1 modification, DNA digestion, signal-on oxidoreduction, and current peak monitoring. An exquisitely designed 5′end-G-rich oligodeoxynucleotide was synthesized bearing a methylated cytosine (5-mC), which formed into hairpin dsDNA with the MspI recognition sequence (C m C GG/GGCC). Then hairpin dsDNA was fabricated onto gold nanoparticles modified glassy carbon electrode (DNA/AuNPs/GCE) via Au-S bond. The combination uses of restriction enzyme MspI and hemin converted fabricated-dsDNA into peroxidase-mimicking DNAzyme, thereby promoting the reduction of H 2 O 2 with a current peak. However, the current peak was extremely decreased once TET1 and T4 β-GT were used in advance. We confirmed a delicately linear relationship matching between the current difference and TET1 activity from 0.7 to 10.5 ng μL −1 with a detection limit of 0.027 ng μL −1 , which outcompeted the former methods at least one order of magnitudes. The TET1 activity evaluation in the existence of Bobcat339 was also tested as the proof of concept of inhibitors screening. Our strategy provides a novel, label-free, and sensitive electrochemical approach that enables us to complete both TET1 activity evaluation and potential TET1 inhibitors screening.
研究领域:
生物化学
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分子生物学
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分析化学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）