ANALYTICA CHIMICA ACTA
·
2026
·
IF 6.1
Huiyan Cao,Xi Cheng,Li Huang,Haotian Zhang,Ziyi Liu,Wenjun Yang,Song-Mei Liu,Fang Wang
Background N6-methyladenosine (m6A), the most abundant internal modification of eukaryotic mRNA, post-transcriptionally regulates gene expression, and its dysregulation is intimately linked to tumorigenesis and multiple human diseases. However, existed detection methods including high-performance liquid chromatography (HPLC) suffer from obvious drawbacks such as site specificity constraints, high cost, reliance on sophisticated instruments and tedious operation, making them unsuitable for rapid on-site detection. Therefore, the development of a novel, rapid, on-site and cost-effective detection strategy for m6A is of great significance and urgent demand. Results This study developed an electrochemical biosensor based on DNA tetrahedrons (TDN) and gold-antibody nanoparticle (AuNPs-ab) for highly sensitive detection of m6A-RNA modifications. After enhancing conductivity through gold nanoparticle modification, TDN was stably immobilized on the electrode surface via Au-S bonds. The rigid structure of TDN was utilized to optimize surface flatness and spatial architecture, enabling precise anchoring of the antibody probe. Target m6A-RNA captured by antibodies undergoes biotin-streptavidin-mediated horseradish peroxidase (HRP) conjugation, catalyzing a hydrogen peroxide–hydroquinone (H 2 O 2 –HQ) redox cycle to generate amplified current signals. The sensor exhibits excellent linear response within the 0.001–100 nM range, and the detection limit is 0.105 pM, demonstrating high specificity and stability (signal retention rate >88% after 15 days). Furthermore, this method has been successfully applied to detect target samples spiked into human serum and total RNA extracted from hepatocellular carcinoma cells. Significance This sensing platform combines the compact size and scalable batch-production advantages of LIG electrodes, offering a novel approach for precise detection of tumor-associated m6A modifications and rational design of disposable, portable point-of-care testing (POCT) devices. It demonstrates significant potential for rapid screening of key epigenetic biomarkers and real-time monitoring across diverse clinical and biological samples.
✓ 已验证
DOI
PubMed
高影响因子
CANCER LETTERS
·
2023
·
IF 9.7
Jiaoshun Chen,Haoxiang Zhang,Chaoyang Xiu,Chenggang Gao,Shihong Wu,Jianwei Bai,Qiang Shen,Tao Yin
In eukaryotes, N 6 -methyladenosine (m6A) is the most prevalent epigenetic alteration. Methyltransferase-like 3 (METTL3) is a key player in the control of m6A, although its function in pancreatic cancer is incompletely understood. In this study, we examined the role that METTL3 plays in pancreatic cancer cell proliferation and stemness. We discovered that in pancreatic cancer cells, METTL3-mediated m6A alterations regulate ID2 as a downstream target. The stability of ID2 mRNA was decreased and m6A modification was effectively eliminated by METTL3 knockdown in pancreatic cancer cells. We also demonstrate that m6a-YTHDF2 is necessary for the METTL3-mediated stabilization of ID2 mRNA. Additionally, we show that ID2 controls the stemness molecules NANOG and SOX2 via the PI3K-AKT pathway to support pancreatic cancer growth and stemness maintenance. Our data suggest that METTL3 may post-transcriptionally upregulate ID2 expression in an m6A-YTHDF2-dependent manner to further promote the stabilization of ID2 mRNA, which may be a new target for pancreatic cancer treatment.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Cell Death & Disease
·
2025
·Vol. 16
·期 1
·1-15
·
IF 9.6
Cen Junjie,Zhao Dongliang,Shi Xin,Chen Jinlong,Zhou Hang,Liang Yanping,Gui Chengpeng,Chen Wei,Luo Junhang,Chen Xu
Renal cell carcinoma (RCC) remains a significant clinical challenge due to its increasing incidence and resistance to conventional therapy. This study identifies an unprecedented mechanism by which mesencephalic astrocyte-derived neurotrophic factor (MANF) contributes to RCC progression. From single-cell transcriptome, we observed a significant upregulation of MANF in RCC compared to its progenitor cells, the proximal tubular cells. Mechanistically, decreased N6-methyladenosine (m6A) modification on MANF mRNA, mediated by the upregulation of the m6A demethylase ALKBH5, led to reduced RNA degradation and increased MANF protein expression. Furthermore, while the common von Hippel-Lindau (VHL) deletion in RCC results in the accumulation of misfolded proteins and endoplasmic reticulum (ER) stress, increased MANF efficiently mitigated ER stress by binding to phosphorylated inositol-requiring enzyme-1 alpha (IRE1α) and inhibited its phosphorylation, indicating that MANF-mediated ER stress resistance compensates for the negative effects of VHL depletion and contributes to the survival of RCC. Our findings highlight a potential therapeutic strategy for RCC by targeting the m6A-mediated regulation, directly degrading MANF protein or inhibiting its function.
研究领域:
分子生物学
·
癌症研究
·
生物化学
·
细胞生物学
·
转化医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2024
·Vol. 29
·期 19
·4718
·
IF 4.2
Jingyi Guo,Mei Zhao,Xia Kuang,Zilin Chen,Fang Wang
Laser-induced graphene (LIG) possesses characteristics of easy handling, miniaturization, and unique electrical properties. We modified the surface of LIG by electropolymerizing β-cyclodextrin (β-CD), which was used to immobilize antibodies on the electrode surface for highly sensitive detection of targets.N6-methyladenosine (m6A) is the most prevalent reversible modification in mammalian messenger RNA and noncoding RNA, influencing the development of various cancers. Here, β-CD was electropolymerized to immobilize the anti-m6A antibody, which subsequently recognized the target m6A. This was integrated into the catalytic hydrogen peroxide–hydroquinone (H2O2-HQ) redox system using phos-tag-biotin to generate electrochemical signals from streptavidin-modified horseradish peroxidase (SA-HRP). Under optimal conditions, the biosensor exhibited a linear range from 0.1 to 100 nM with a minimum detection limit of 96 pM. The method was successfully applied to the recovery analysis of m6A from HeLa cells through spiking experiments and aims to inspire strategies for point-of-care testing (POCT).
研究领域:
生物化学
·
分析化学
·
生物医学工程
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
International Journal of Medical Sciences
·
2024
·
IF 3.2
Mi Tang,Min Wang,Zhirong Wang,Bo Jiang
Aortic aneurysm and dissection (AD) represent a critical cardiovascular emergency with an alarmingly high mortality rate. Recent research has spotlighted the overexpression of genes associated with the m6A modification in AD patients, linking them to the presence of inflammatory M1-type macrophages. Moreover, glycolysis is widely recognized as a key feature of inflammatory M1-type macrophages, but biomarkers linking glycolysis and macrophage function to promote disease progression in AD have not been reported. We conducted an analysis of aortic immune cell infiltration, macrophages, and m6A-related biomarkers in AD patients using bioinformatics techniques. Subsequently, we employed a combination of RT-PCR, WB, and immunofluorescence assays to elucidate the alterations in the expression of M1- and M2-type macrophages, as well as markers of glycolysis, following the overexpression of key biomarkers. These findings were further validated in vivo through the creation of a rat model of AD with knockdown of the aforementioned key biomarkers. The findings revealed that the m6A-modified related gene RBM15 exhibited heightened expression in AD samples and was correlated with macrophage polarization. Upon overexpression of RBM15 in macrophages, there was an observed increase in the expression of M1-type macrophage markers CXCL9 and CXCL10, alongside a decrease in the expression of M2-type macrophage markers CCL13 and MRC1. Furthermore, there was an elevation in the expression of glycolytic enzymes GLUT1 and Hexokinase, as well as HIF1α, GAPDH, and PFKFB3 after RBM15 overexpression. Moreover, in vivo knockdown of RBM15 led to an amelioration of aortic aneurysm in the rat AD model. This knockdown also resulted in a reduction of the M1-type macrophage marker iNOS, while significantly increasing the expression of the M2-type macrophage marker CD206. In conclusion, our findings demonstrate that RBM15 upregulates glycolysis in macrophages, thus contributing to the progression of AD through the promotion of M1-type macrophage polarization. Conversely, downregulation of RBM15 suppresses M1-type macrophage polarization, thereby decelerating the advancement of AD. These results unveil potential novel targets for the treatment of AD.
研究领域:
心血管疾病
·
免疫学
·
分子生物学
·
生物信息学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Integrative Agriculture
·
2026
·
IF 5.7
Zimeng Xin,Shun Chen,Ruiti Ren,Jiaqi Liu,Shizhe Xiang,Yuchu Han,Liangpeng Ge,Yongming Zhou,Jing Sun,Xinxia Wang
The gut microbiota plays a critical role in regulating host fat deposition, yet the underlying mechanisms remain poorly understood. To investigate this question, this study employed germ-free (GF) and fecal microbiota transplantation (FMT) piglet models to systematically elucidate how microbiota-derived metabolites modulate fat deposition via N 6 -methyladenosine (m 6 A) RNA methylation. Although the FMT group showed no significant change in body weight compared with the GF group, it exhibited markedly increased subcutaneous and intramuscular fat deposition, as evidenced by larger adipocyte size ( P <0.001), elevated serum and longissimus dorsi muscle triglyceride levels ( P <0.05), and upregulated expression of adipogenic genes ( P < 0.05). Non-targeted metabolomics revealed elevated levels of bile acids, including hyodeoxycholic acid (HDCA), 12-ketodeoxycholic acid (12KDCA), and 3b-hydroxy-5-cholenoic acid (3b-h5-CA), as well as tryptophan metabolites indoxyl sulfate (IS) and L-kynurenine (L-Kyn) in the FMT group, and these metabolites were significantly correlated with lipid metabolism parameters. Mechanistic studies showed that these metabolites reduced m 6 A modification levels in mouse stromal vascular fraction (SVF) cells and fibro-adipogenic progenitor (FAPs) cells by upregulating fat mass and obesity-associated protein (FTO) and downregulating methyltransferase-like 3 (METTL3) ( P < 0.05), consistent with the in vivo observations in piglets. Furthermore, Bacteroides , Lactobacillus , Parabacteroides , and Akkermansia were identified as key genera involved in bile acid and tryptophan metabolism. Together, these findings reveal a gut microbiota–metabolite– m 6 A regulatory axis in fat deposition in pigs, providing new insights into host–microbiota interactions and offering potential strategies for improving metabolic health and meat quality in livestock.
✓ 已验证
DOI
高影响因子
BIOCHEMICAL PHARMACOLOGY
·
2026
·
IF 6.5
Enjia Zhou,Zeyu Xing,Mingchao Wang,Dongxu Lu,Mengyao Hao,Guodong Tang,Shuying Li,Yiran Chen,Wenjing Zhang,Xinyu Li,Bolin Zhang,Fang Wang,Wenhao Ju,Jing Jin
Epigenetic modifications, particularly N6-methyladenosine (m6A) modifications, alter the phenotype of macrophages. Among the m6A-related proteins, methyltransferase 3 (METTL3), as the catalytic core methyltransferase subunit, plays an essential role in regulating the function of macrophages. This study aimed to explore novel m6A inhibitors from natural products that target METTL3 to modulate the phagocytic function of macrophages in the treatment of colon cancer. In this work, first, a high-throughput screening of biological m6A by colorimetric assay was conducted to rapidly identify candidate inhibitors from a natural product library. Second, bioinformatics analysis and molecular biological methods revealed the underlying mechanisms through which METTL3 regulates macrophage-mediated phagocytosis. Subsequently, Mettl3 f/f Lyz2 -Cre mice were generated to further elucidate the mechanism through which METTL3 inhibition promotes the phagocytic function of macrophages by modulating the mRNA stability of SIRPA, a key phagocytic-related protein, in vivo . We determined that chlorogenic acid (CHA) is a potential therapeutic METTL3 inhibitor because of its influence on METTL3 enzymatic activity and expression. METTL3 inhibition promoted the phagocytic function of macrophages. METTL3 increases the mRNA stability of the intrinsic immune checkpoint SIRPA by activating IGF2BP2 in an m6A-dependent manner, which subsequently increases its protein expression and suppresses the phagocytosis of macrophages both in vivo and in vitro . In conclusion, CHA is a candidate inhibitor compound that targets the METTL3–SIRPA axis through enhancing the phagocytic function of macrophages and is thus a promising therapy for treating colon cancer.
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2026
·
IF 10.6
Yan Zhang,Fuqing Yi,Ying He,Yidan Ma,Xinyi Mu,Yanqing Geng,Xin Yin,Rufei Gao,Liping Yan,Xuemei Chen,Junlin He
Dibutyl phthalate (DBP), a common environmental endocrine-disrupting chemical, is known to impair female reproduction, yet its specific effects on ovarian reserve and underlying epigenetic mechanisms are not fully understood. Here, using gestational exposure models ( in vivo ) and fetal ovarian culture models ( in vitro ), we show that gestational DBP exposure disrupts the ovarian reserve through a dual assault: impairing primordial follicle formation ("input") and, more critically, driving primordial follicle premature activation ("output"). Mechanistically, DBP induces METTL14-dependent m⁶A hypermethylation, which activates the PI3K-AKT-FOXO3a signaling axis to promote aberrant follicular activation. Knockdown of Mettl14 rescues this pathway activation and the follicular disruption. Long‑term follow‑up revealed that such precocious activation leads to progressive ovarian reserve depletion, culminating in a premature ovarian insufficiency (POI)-like phenotype with hormonal dysfunction, reduced oocyte quality, and compromised fertility in adult offspring. Collectively, these findings indicate that gestational DBP exposure activates the PI3K-AKT-FOXO3a signaling pathway via METTL14-mediated m⁶A modification, causing impaired follicle formation, follicular over-activation, and ultimately ovarian reserve depletion and a POI-like phenotype in offspring. This study reveals a novel m⁶A‑mediated epigenetic mechanism in plasticizer-induced ovarian toxicity, advancing our understanding of POI etiology and highlighting a significant environmental risk to female fertility.
✓ 已验证
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2021
·
IF 6.291
Luyao Li,Mei Zhou,Biyun Chen,Qin Wang,Shuya Pan,Yaxuan Hou,Jiamin Xia,Xue Zhou
Cadmium is a carcinogenic heavy metal that poses a severe threat to human beings. The underlying mechanism, however, remains elusive. N6-methyladenosine (m6A) is the most abundant post-transcriptional modification in mRNA that regulates RNA metabolism. Emerging evidence shows that m6A is involved in the pathogenesis of various cancers. In this study, human bronchial epithelial BEAS-2B cells were transformed by exposing to 2 μM of cadmium for 20 weeks to investigate the role of m6A in cadmium carcinogenesis. We found the level of m6A in mRNA was significantly decreased in cadmium-transformed BEAS-2B cells, and this change was regulated by m6A demethylase ALKBH5. ALKBH5 was significantly upregulated in the middle and late stages of cell transformation at week 8, 12, 16 and 20. Knockdown of ALKBH5 in cadmium-transformed cells alleviated cell proliferation, migration, invasion, and anchorage-independent growth, but co-transfection with ALKBH5 siRNA and PTEN siRNA restored the inhibitory effects of ALKBH5 knockdown on those transformation properties. ALKBH5 decreased the m6A level of PTEN mRNA, resulting in its instability and reduction of PTEN protein expression. These results indicate that ALKBH5-mediated demethylation m6A at PTEN mRNA is involved in cadmium-induced cell transformation. Our study provides a new perspective for the involvement of m6A modification in cadmium carcinogenesis.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CELLULAR AND MOLECULAR LIFE SCIENCES
·
2026
·
IF 6.5
Qu Qiao,Li Zhilong,Wang Dalu,Wu Di,Yin Hongzhuan
Colorectal cancer (CRC) remains a major contributor to cancer-related mortality worldwide. Although minichromosome maintenance protein 10 (MCM10) is known for its role in DNA replication, its involvement in CRC progression has not been fully elucidated. To identify key oncogenic drivers, RNA sequencing was performed on seven paired CRC and adjacent normal tissues, revealing significant upregulation of MCM10 (Log2FC = 2.359, P = 0.003). Public GEO datasets (GSE240623, GSE200427) further confirmed MCM10 overexpression in CRC tissues. This finding was validated in 40 paired clinical samples using qPCR and Western blotting. Prognostic relevance was assessed using hazard ratio (HR). Functional assays in vitro and in vivo were conducted to assess the effects of MCM10 on proliferation, migration, invasion, and M2 macrophage polarization. The m6A modification of MCM10 mRNA was evaluated through METTL3 and YTHDF1 interaction analysis in HCT116 cells. MCM10 was significantly upregulated in CRC tissues and associated with poor prognosis (HR = 1.54, P = 0.00077). MCM10 overexpression promoted CRC cell proliferation and invasion and was correlated with elevated M2 macrophage infiltration in tumor xenografts (6.16 ± 0.85% vs. 11.7 ± 1.13%, P < 0.05). Conditioned media from MCM10-overexpressing cells enhanced THP-1 macrophage polarization by increasing secretion of CCL2, CCL5, and IL10. Mechanistically, METTL3-mediated m6A modification stabilized MCM10 mRNA in a YTHDF1-dependent manner. MCM10 is aberrantly overexpressed in CRC and linked to worse clinical outcomes. Its oncogenic effects involve METTL3/YTHDF1-mediated mRNA stabilization and enhanced M2 macrophage polarization, suggesting a potential role in immune microenvironment modulation.
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF GASTROENTEROLOGY
·
2025
·
IF 5.5
Ren Bingyu,Li Han,Liu Shenglu,Huang Zhiwei,Bai Junjie,Jiang Zhonghao,Xu Tongjie,Gu Boyuan,Yu Wenhao,Sun Lei,Tan Peng,Fu Wenguang
Background and Aims Inflammatory cell infiltration in the liver is a hallmark of metabolic dysfunction-associated fatty liver disease (MAFLD). However, the pathological events that trigger the infiltration of inflammatory cells to mediate MAFLD pathogenesis remains poorly understood. This study aims to investigate the function and mechanism of Hic-5 on hepatic inflammation of MAFLD. Methods MAFLD animal models were fed a methionine- and choline-deficient (MCD) diet in Hic-5 knockout mice. Liver tissues were analyzed by immunohistochemical staining, immunofluorescence and flow cytometry, with a particular focus on the impact on the immune microenvironment. Results Hic-5 deficiency alleviates the severity of MAFLD, particularly the inflammation response. Gain- and loss-of-function experiments revealed that Hic-5 deficiency results in decreased neutrophil proliferation and increased apoptosis, as well as impaired migration. Conversely, Hic-5 overexpression had the opposite effects. This study confirmed that METTL3-mediated methylation of m 6 A stabilizes Hic-5 mRNA and promotes its expression, which in turn regulates the infiltration of neutrophils by the CXCL1-CXCR2 axis. Conclusions The study reveals the role of Hic-5 in regulating neutrophils and indicates that it may be a potential therapeutic target for MAFLD.
研究领域:
肝病学
·
免疫学
·
分子生物学
·
病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BRITISH JOURNAL OF PHARMACOLOGY
·
2026
·
IF 7.5
Yufen Xu,Xiaoli Tan,Xiaodong Lv,Qi Yang,Xingxing Ke,Wenyu Chen
Background and Purpose IGF2BP1 stabilizes oncogenic mRNAs via m6A binding to drive lung adenocarcinoma (LUAD) progression, yet how this modification orchestrates immune evasion remains unclear. Experimental Approach Bioinformatics, RT-qPCR and Western Blotting (WB) first profiled IGF2BP1 expression in LUAD. RNA pull-down, IL-11 stability assays, ELISA and confocal imaging screened and validated IGF2BP1/IL-11 as the key oncogenic axis. ELISA, cytotoxicity experiments and flow cytometry were applied in the exploration of the function of the IGF2BP1/IL-11 axis in LUAD. Bioinformatics analysis, IGF2BP1 half-life detection, WB and ubiquitination experiments were employed to identify the interaction between KCTD10 and IGF2BP1. Allograft experiments were conducted to validate the impact of the KCTD10/IGF2BP1/IL-11 axis on tumour immune escape. Clinical correlation analysis was based on samples of LUAD patients before and after PD-1/PD-L1 inhibitor treatment. Key Results IGF2BP1 was up-regulated in LUAD, exhibiting an association with adverse prognosis. Mechanistically, we observed that IGF2BP1 stabilized IL-11 mRNA in an N6-methyladenosine (m6A)-dependent manner, leading to higher protein expression of IL-11, thereby dampening the cytotoxicity and proliferation of CD8 + cells as well as the expression of TNF-α, IFN-γ and IL-2. Furthermore, KCTD10 overexpression facilitated ubiquitination and degradation of IGF2BP1, and its combination with an anti-IL-11 monoclonal antibody dramatically enhanced therapeutic efficacy. Conclusion and Implications These findings reveal that KCTD10 degrades IGF2BP1 by ubiquitination, regulates m6A modification of IL-11, and restrains immune evasion in LUAD. Our study identifies potential targets for immunotherapy and proffers new therapeutic options for LUAD patients.
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）