ACTA PHARMACOLOGICA SINICA
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2026
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IF 10.4
Xu Lu,Ma Yan,Yao Wang,Qiu Ya-ju,Xu Chao,Xu Fang-yue,Zou Zhi-miao,Dai Wu-min,Gong Wan-gang,Ma De-ning,Chen Xiao-wan,Ye Wen-kai,Huang Yi-yun,Chen Hai-tao,Hong Lue,Mo Jia-ping,Feng Jian-guo,Yu En-yan,Chen Wei,Zhang Yu-hua,Yao Qing-hua,Li Xia
5-Fluorouracil (5-FU) resistance in colorectal cancer (CRC) involves oxidative stress mechanisms, but the role of epitranscriptomic regulation remains unclear. This study investigates how oxidative post-translational modifications of the N6-methyladenosine (m 6 A) methyltransferase METTL16 contribute to 5-FU resistance. Parental (HCT8, HCT15) and 5-FU-resistant CRC cells were compared using redox proteomics, m 6 A-seq, RNA-seq, and functional assays. METTL16 S-glutathionylation was assessed via streptavidin pulldown and mass spectrometry. IGF2BP3’s role was validated through knockdown/overexpression, patient-derived organoids (PDOs), and xenograft models. Clinical relevance was evaluated in 112 CRC patient tissues and data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Drug synergy was tested using isoliquiritigenin (ISO) combined with 5-FU. Resistant cells exhibited elevated global protein S-glutathionylation and reduced m 6 A RNA methylation. METTL16 underwent site-specific S-glutathionylation at Cys548, leading to its degradation and subsequent m 6 A loss. Integrative omics identified IGF2BP3 as a key METTL16 target: reduced m 6 A on IGF2BP3 mRNA enhanced its nuclear export, splicing, and overexpression. IGF2BP3 knockdown sensitized resistant cells to 5-FU in vitro and in vivo, while overexpression conferred resistance. Mechanistically, IGF2BP3 destabilized CFTR mRNA (an ABC transporter), reducing 5-FU uptake. Clinically, high IGF2BP3 correlated with poor survival and 5-FU resistance in CRC patients. Targeting IGF2BP3 with ISO synergized with 5-FU, overcoming resistance in cells and xenografts. Oxidative stress-induced METTL16 S-glutathionylation drives 5-FU resistance by reducing m 6 A modification, enabling IGF2BP3 overexpression and CFTR suppression. IGF2BP3 is a biomarker of clinical resistance, and its targeting with ISO represents a promising combinatorial strategy to restore 5-FU efficacy in CRC.
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ANALYTICA CHIMICA ACTA
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2026
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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.
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International Journal of Medical Sciences
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2024
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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
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免疫学
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分子生物学
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生物信息学
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Biochimica et Biophysica Acta-Gene Regulatory Mechanisms
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2021
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IF 4.49
Da Cheng,Cichun Wu,Ying Li,Yao Liu,Juan Mo,Lei Fu,Shifang Peng
Hepatitis B virus (HBV)-associated acute-on-chronic liver failure (ACLF) is a clinical syndrome of severe liver damage. HBV infection is affected by N6-methyladenosine (m 6 A) RNA modification. Here, we investigated whether methyltransferase-like 3 (METTL3)-mediated m 6 A methylation can affect ACLF. Human hepatic cells (THLE-2) were treated with lipopolysaccharide (LPS) to induce cell damage. Proliferation, apoptosis and m 6 A modification were measured by MTT assay , flow cytometry and Dot blot assay. Our results showed that HBV infection significantly enhanced the levels of m 6 A modification and elevated the expression of METTL3 and mature-miR-146a-5p in THLE-2 cells, which was repressed by cycloleucine (m 6 A inhibitor). METTL3 overexpression enhanced m 6 A modification and promoted mature-miR-146a-5p expression. METTL3 overexpression promoted HBV replication and apoptosis, enhanced the levels of pro-inflammatory cytokines, hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg), and repressed cell proliferation in THLE-2 cells, which attributed to repress miR-146a-5p maturation. Moreover, a severe liver failure mouse model was established by HBV infection to verify the impact of METTL3 knockdown on liver damage in vivo . HBV-infection led to a severe liver damage and increase of apoptosis in hepatic tissues of mice, which was abolished by METTL3 knockdown. METTL3 knockdown reduced METTL3 expression and impeded miR-146a-5p maturation in HBV-infected mice. In conclusion, this work demonstrates that METTL3 inhibition ameliorates liver damage in mouse with HBV-associated ACLF, which contributes to repress miR-146a-5p maturation. Thus, this article suggests a novel therapeutic avenue to prevent and treat HBV-associated ACLF.
研究领域:
分子生物学
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肝病学
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病毒学
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药理学
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JOURNAL OF GASTROENTEROLOGY
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2025
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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
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免疫学
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分子生物学
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病理学
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CELLULAR AND MOLECULAR LIFE SCIENCES
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2026
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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.
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Journal of Integrative Agriculture
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2026
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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.
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BIOCHEMICAL PHARMACOLOGY
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2026
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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.
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JOURNAL OF HAZARDOUS MATERIALS
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2026
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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.
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SENSORS AND ACTUATORS B-CHEMICAL
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2023
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IF 8.4
Xinghui Zhou,Chengpeng Li,Wanxue Wang,Feng Zhang,Yi Zhou,Danping Chen,Zhenchao Wang
N 6 -methyladenosine (m 6 A) is a crucial post-transcriptional epigenetic modification in RNAs which associates with numerous cancers development. Here, a cost-effective, and easy-to-operate fluorescence strategy for m 6 A analysis was developed based on the different cleavage efficiency of 8–17 DNAzyme for methylation and unmethylation sites. By arranging 8–17 DNAzyme to recognize and cleave 5'-rAG-3' and 5'-rm 6 AG-3' motifs in the target sequence respectively, various short ssDNA oligoes were produced, leaving different amounts of target sequence. Benefiting from the varying degrees of retardation of CHA amplification reaction, the tiny discrepancy between m 6 A and unmethylated adenine (A) can be distinguished. Moreover, we found that 8–17 DNAzyme was more sensitive to m 6 A than to A. Importantly, the strategy we proposed not only avoids sophisticated chemical labelling and expensive proteins, but also can be easily performed on the ordinary heating apparatus of accessible laboratory. The DCHA strategy demonstrated good analytical performance in complex biological matrices, and was successfully applied to hepatocellular carcinoma-related m 6 A analysis in CTNNB1, MGAT5 and SOCS-2 mRNAs with satisfactory results, showing its potential in clinical analysis.
研究领域:
分子生物学
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生物化学
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癌症生物学
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表观遗传学
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BRITISH JOURNAL OF PHARMACOLOGY
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2026
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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.
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LABORATORY INVESTIGATION
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2024
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IF 5.1
Rui Hou,Yadong Wang,Shiyao Cao,Xinrui Sun,Luo Jiang
High-grade serous ovarian cancer (HGSOC) remains the most lethal female cancer by far. Herein, clinical HGSOC samples had higher N 6 -methyladenosine (m 6 A) modification than normal ovarian tissue, and its dysregulation had been reported to drive aberrant transcription and translation programs. However, Kringle containing transmembrane protein 2 (KREMEN2) and its m 6 A modification have not been fully elucidated in HGSOC. In this study, the data from the high-throughput mRNA sequencing (RNA-seq) of clinical samples were processed using the weighted correlation network analysis (WGCNA) and functional enrichment analysis. Results revealed that KREMEN2 was a driver gene in the tumorigenesis of HGSOC and a potential target of m 6 A demethylase fat-mass and obesity-associated protein (FTO). KREMEN2 and FTO levels were upregulated and downregulated, respectively, and correlation analysis showed a significant negative correlation in HGSOC samples. Importantly, upregulated KREMEN2 was remarkably associated with lymph node (LN) metastasis, distant metastasis, peritoneal metastasis, and high FIGO stage (Ⅲ/Ⅳ), independent of the age of patients. KREMEN2 promoted the growth of HGSOC in vitro and in vivo , which was dependent on FTO. The methylated RNA immunoprecipitation-qPCR and RNA immunoprecipitation assays were performed to verify the m 6 A level and sites of KREMEN2. FTO overexpression significantly decreased m 6 A modification in 3'UTR and 5'UTR regions of KREMEN2 mRNA and downregulated its expression. In addition, we found that FTO-mediated m 6 A modification of KREMEN2 mRNA was recognized and stabilized by the m 6 A reader IGF2BP1 rather than IGF2BP2 or IGF2BP3. This study highlights the m 6 A modification of KREMEN2 and extends the importance of RNA epigenetics in HGSOC.
研究领域:
癌症生物学
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分子生物学
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表观遗传学
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肿瘤学
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