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N6-methyladenosine(m6A)is the most prevalent modification that regulates gene expression in eukaryotes.It regulates splicing,degradation,stability,and translation of RNA.Numerous studies have demonstrated the close association between m6A methylation and tumor development,highlighting its crucial role in regulating tumor immune response.The m6A modification actively participates in governing immune cell differentiation and maturation as well as modulating anti-tumor immune responses.Within the tumor microenvironment,m6A modification can also impact the recruitment,activation,and polarization of immune cells,thereby either promoting or inhibiting tumor cell proliferation and metastasis.Consequently,it plays a pivotal role in reshaping the tumor immune microenvironment.In recent years,immunotherapy for tumors has been increasingly applied to clinical practice with notable success achieved through approaches such as immune checkpoint inhibitor therapy and adoptive cell immunotherapy.Targeting m6A modifications to interfere with the immune system,such as targeting dysregulated m6A regulators through small molecule inhibitors and inducing immune cell reprogramming,can improve anti-tumor immune response and strengthen immune cells' ability to recognize and kill tumor cells.The m6A modification represents a novel avenue for potential clinical application within tumor immunotherapy.This review provides a comprehensive summary of the regulatory impact of m6A methylation modification on immune cells in the context of cancer,while also delving into novel targets for tumor immunotherapy.
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Gastric cancer(GC)is one of the most common malignancies in the digestive system.Many patients are found in advanced stage and have a poor prognosis.Surgery and chemotherapy remain the main treatments for gastric cancer.N6-methyladenosine(m6A)is a hot topic in tumor research in recent years.As the most common form of RNA modification in eukaryotes,m6A can regulate various stages of the RNA cycle,including RNA splicing,processing,degradation,and translation,thereby regulating RNA expression and function,playing a critical role in various pathways such as cell differentiation,development,and metabolism.The m6A demethylase can remove methyl groups on RNA,ensuring that m6A methylation is a dynamic and reversible process.As a key enzyme in the m6A methylation process,the imbalance of m6A demethylases fat mass and obesity-associated protein(FTO),AlkB homolog 5(ALKBH5)and ALKBH3 regulate the progression of gastric cancer through various mechanisms,which is closely related to the occurrence and development of gastric cancer.These m6A demethylases regulate the signaling pathway,alter the proliferation and invasion ability of gastric cancer cells,affect its resistance to chemotherapy drugs,participate in regulating the immune response and mitochondrial metabolism of gastric cancer,and affect the growth of gastric cancer cells.They are expected to become a novel therapeutic target.This article comprehensively summarizes the molecular mechanism of m6A demethylase involved in the occurrence and development of gastric cancer,and the relationship between its expression and function,and biological characteristics of m6A demethylase were reviewed,aiming to provide new research ideas for early diagnosis and targeted treatment of gastric cancer.
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Nonalcoholic fatty liver disease(NAFLD)is a metabolic liver disease that ranges from relatively benign hepatic steatosis to nonalcoholic steatohepatitis(NASH).NASH is characterized by persistent liver damage,inflammation,and fibrosis which significantly increases the risk of end-stage liver diseases,such as liver cirrhosis and hepatocellular carcinoma.The pathogenesis of NAFLD/NASH is not yet fully understood,but its recent epigenetic advances have provided new insights into the mechanisms of this disease.This review summarized recent progress in this area which has laid a solid foundation for elucidating the pathogenesis of NAFLD and provides potential targets for early detection,diagnosis,and treatment of this disease.
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Acute kidney injury(AKI)is a global public health problem with high morbidity,high mortality and costly treatment cost.The pathogenesis of AKI is very complex,and the treatment strategies for AKI are lim-ited,then it is very matter to explore the pathophysiological mechanism and potential therapeutic targets of acute kidney injury.N6-methyladenosine(m6A)is the most abundant and extremely conservative epigenetic modification in eukaryotic,which is a dynamic and reversible process involving in splicing,nuclear export,translation,stabil-ity,and higher structure of RNA,and regulated by three regulatory factors:methyltransferase,demethylase and methylated reading protein.Current studies have found that m6A plays an important regulatory role in AKI and can be a potential therapeutic target for AKI.In this review,we provide a brief description of m6A and summarize the impact of m6A on AKI and possible future study directions for this research.
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Post-stroke cognitive impairment(PSCI)is mainly manifested as learning and memory disorders.Highly enriched RNA m6A methylation modification in mammalian brain is involved in glial cell-mediated neuroinflammation.Given that neuroinflammation is the main mechanism for neural damage and spatial and memory impairment of PSCI,it is speculated that RNA m6A methylation modification can regulate the inflammatory response of glial cells after stroke to improve PSCI.This review summarizes and analyzes the role of RNA m6A methylation modification in the development of PSCI and analyzes its detailed mechanism of regulating glial cell-mediated inflammation,which will provide reference for researchers in this field.
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BACKGROUND:It is known that N6-methyladenosine(m6A)plays a role in the pathogenesis of various diseases and studies have suggested its involvement in the pathologic changes of steroid-induced femoral head necrosis(SNFH).However,research on m6A methylation modifications in steroid-induced femoral head necrosis is limited. OBJECTIVE:Using bioinformatics methods to identify the differential expression of m6A-related genes in steroid-induced femoral head necrosis and to predict miRNAs associated with these genes to further elucidate the role and mechanism of m6A methylation in steroid-induced femoral head necrosis. METHODS:Differential gene expression between steroid-induced femoral head necrosis and control groups was analyzed using GSE123568 gene expression data and identified using the"limma"package in R.Functional enrichment analysis was performed on the differentially expressed genes.Differential analysis of the related genes was carried out using the"ggstatsplot"package in R.The differential genes were cross-validated using the GSE74089 dataset.An mRNA-miRNA regulatory network was constructed,and co-expression analysis was performed on the module genes followed by enrichment analysis.Differences in immune cell infiltration between steroid-induced femoral head necrosis and control groups were quantified using the ssGSEA method. RESULTS AND CONCLUSION:Correlation analysis revealed 13 m6A-related genes,and further analysis through the protein-protein interaction network identification and receiver operating characteristic curve analysis showed that YTHDF2 was expected to be a core differential gene as a potential early biomarker.Enrichment analysis indicated that differentially expressed genes were mainly involved in inflammation and immune response and were closely related to osteoclasts.Cross-validation analysis showed that differential gene expression results between the two datasets were consistent.mRNA-miRNA regulatory network analysis revealed that YTHDF2 was negatively correlated with miRNA-27a.Immune infiltration analysis revealed an increase in immune cell infiltration in steroid-induced femoral head necrosis,and YTHDF2 was positively correlated with the infiltration of CD4+T cells.To conclude,m6A-related gene YTHDF2 can serve as a potential biomarker of steroid-induced femoral head necrosis and is valuable for the early clinical diagnosis and treatment of steroid-induced femoral head necrosis.The negative correlation between YTHDF2 and mir-27a and the positive correlation between YTHDF2 and CD4+T cell infiltration provide new insights into the early diagnosis and treatment of steroid-induced femoral head necrosis and shed light on the mechanism of m6A in steroid-induced femoral head necrosis.
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BACKGROUND:The pathogenesis of osteoporosis is complex,and its essence is the weakening of bone formation and the enhancement of bone absorption caused by various reasons,resulting in the imbalance of bone metabolism.In recent years,N6-methyladenosine has been found(N6-methyladenosine,m6A)methylation can prevent and treat osteoporosis by regulating bone metabolism. OBJECTIVE:Taking the regulation of bone metabolism by m6A methylation as an entry point,to systematically sort out and summarize the research progress of m6A methylation in osteoporosis,so as to provide certain theoretical reference bases for the search of new therapeutic targets for osteoporosis. METHODS:CNKI,WanFang,VIP,PubMed,MEDLINE,Nature,and Cochrane databases were retrieved for relevant literature published from database inception to 2023.The keywords were"osteoporosis,m6A methylation,bone metabolism,bone marrow mesenchymal stem cells,osteoblasts,osteoclasts"in Chinese and English.Duplicates and obsolete non-referenced documents were excluded,and a total of 73 standard papers were included for further review. RESULTS AND CONCLUSION:m6A methylation can affect the activity and differentiation of bone marrow mesenchymal stem cells,osteoblasts,and osteoclasts through various pathways to regulate bone metabolism and prevent osteoporosis.The regulatory process of m6A methylation is extremely complex,and its related proteins play different roles in different cells.Even in the same kind of cells,the same type of proteins may have radically different roles,regulating different physiological and pathological processes.
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BACKGROUND:m6A modification has been confirmed to play an important role in the occurrence and development of osteonecrosis of the femoral head;however,the role of m6A modification patterns in steroid-induced osteonecrosis of the femoral head remains unknown. OBJECTIVE:Bioinformatics analysis was performed based on the Gene Expression Omnibus(GEO)database to analyze the differential expression of the m6A gene in steroid-induced osteonecrosis of the femoral head,predict the downstream targeted miRNAs,and investigate the potential pathogenesis. METHODS:Expressing profiles of mRNA data of steroid-induced osteonecrosis of the femoral head were downloaded from GEO database(GSE123568).Differentially expressed genes(DEGs),Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using the R software.After obtaining these differentially methylated m6A genes(m6A-DEGs),we analyzed GO function and KEGG pathway enrichment and compared the correlation among the m6A-DEGs typing according to gene expression.The protein-protein interaction network and core gene subnetwork of m6A-DEGs were constructed using Cytoscape software.The m6A-DEGs-associated potential miRNAs were predicted using the TargetScan,miRTarBase,and miRBD databases.Simultaneously,ChIPBase and hTFtarget databases were used to predict potential transcription factors of seven core genes,then m6A-miRNA and transcription factor-m6A regulatory networks were constructed separately.Finally,the expression levels of the seven core m6A-DEGs were verified by using the GSE74089 dataset. RESULTS AND CONCLUSION:(1)A total of 2 460 common DEGs were screened out from datasets,among which 1 455 genes were upregulated and 1 005 genes were downregulated.(2)A total of 14 m6A-DEGs were identified in the datasets.Among them,11 m6A-DEGs were up-regulated and 3 m6A-DEGs were down-regulated.Differential gene expression was considered significant for m6A-DEGs in steroid-induced osteonecrosis of the femoral head(P<0.05).Spearman correlation analysis showed a significant correlation between m6A-DEGs.(3)GO and KEGG enrichment analysis showed that m6A-DEGs were mainly enriched in myeloid cell differentiation and development,immune and cytokine receptor activity,osteoclast differentiation,AMPK signaling pathway and interleukin-17 signaling pathway.(4)The seven core genes of m6A-DEGs contained YTHDF3,YTHDF1,YTHDF2,ALKBH5,METTL3,HNRNPA2B1,and HNRNPC.A total of 44 miRNAs overlapping were detected in the miRTarBase,miRDB,and TargetScan databases.Totally 79 transcription factors overlapping were found in the ChIPBase and hTFtarget databases.(5)The expression levels of six core m6A-DEGs in the GSE74089 dataset were consistent with those in the GSE123568 dataset.(6)These findings confirm that the seven m6A-DEGs identified through bioinformatics techniques play a regulatory role in the expression of various miRNAs,transcription factors,AMPK,and interleukin-17 signaling pathways,and these genes have a significant impact on the differentiation and development of bone marrow cells as well as osteoclast differentiation in steroid-induced osteonecrosis of the femoral head.Consequently,these findings offer data support and establish a research direction for future investigations into the pathogenesis and targeted therapeutic strategies for steroid-induced osteonecrosis of the femoral head.
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Objective:To investigate the alteration of m6A demethylase AlkB homolog 5 (ALKBH5) expression and its impact on form-deprivation myopia (FDM) retina in guinea pigs.Methods:Thirty normal SPF grade 3-week-old tricolor guinea pigs were randomly divided into normal control group and experimental group, with 15 in each group.In the experimental group, the right eyes were covered as FDM group and the left eyes uncovered were set as self-control group.Ocular biometry was performed at one-week intervals from baseline to week 4 of the experiment.Spherical equivalent was detected by streak retinoscopy and axial length was measured by A-scan ultrasonography.Animals were sacrificed after 4 weeks of modeling.The distribution and expression of ALKBH5 protein in the guinea pig retina was detected by immunohistochemical and immunofluorescence staining.Expression of ALKBH5 mRNA and protein in guinea pig retina was detected by real-time fluorescence quantitative PCR and Western blot, respectively.The use of animals in ophthalmic and vision research followed the tenets of Animal Research in Vision and Ophthalmology, and the study was approved by the Ethics Committee of North Sichuan Medical College (No.2023087).Results:At weeks 2, 3, and 4 after myopia induction, diopters and axial lengths were significantly higher in the FDM group than in the normal control group and the self-control group (all at P<0.001). Immunohistochemistry and immunofluorescence assays showed that ALKBH5 protein was expressed in the retinal nerve fiber layer, rod/cone photoreceptor cells, and retinal pigment epithelium (RPE) layer, and was highly expressed in the retinal nerve fiber layer and RPE layer.The relative ALKBH5 immunofluorescence intensity in the normal control group, self-control group and FDM group was 1.000±0.204, 0.874±0.076 and 0.571±0.053, respectively, which was lower in the FDM group than in the normal control and self-control groups, showing statistically significant differences ( t=4.069, P=0.006; t=5.176, P=0.014). After 4 weeks of modeling, ALKBH5 mRNA and protein expressions were significantly lower in FDM group than in normal control and self-control groups (both at P<0.01). Conclusions:The expression of m6A demethylase ALKBH5 is decreased in the retina of FDM guinea pigs, suggesting that ALKBH5 and related m6A methylation modification may be involved in the development and progression of myopia.
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Autophagy is an important mechanism to maintain cellular function and metabolism,whereas ab-normal autophagy can cause the advent and worsening of various diseases.N6-Methyladenosine(m6A)RNA methylation is a reversible RNA modification,which is regulated by m6A methyltransferase,m6A demethylase and m6A-binding protein.Studies have shown that autophagy-related genes promote or attenuate autophagy level dependent on the regulation of m6A,and then participate in the process of diseases.This paper reviews the progress of m6A modification regulatory enzymes and their binding proteins in regulating cell autophagy to provide reference for future researches.
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Objective @# To investigate the correlation between the expression level of YTHDF1 in oral squamous cell carcinoma ( OSCC) and clinicopathologic features and its potential prognostic value．@*Methods @#The expression of YTHDF1 in 132 OSCC tissues and 66 paracancerous tissues was detected by immunohistochemistry (IHC) ，and the expression of YTHDF1 protein in OSCC cell lines was detected by Western blot．The correlation between YTHDF1 and clinicopathological features was analyzed by chi-square test．Kaplan-Meier and Cox factors were used to analyze the factors affecting the survival time of the patients and draw the survival curves of the YTHDF1 gene to evaluate its potential clinical significance. @*Results @#The expression of YTHDF1 in OSCC tissues was higher than that in para- cancerous tissues (P＜0. 001) ，and the expression of YTHDF1 protein increased in OSCC cell lines compared with normal oral epithelial keratinocytes (P ＜0. 001) ．The expression of YTHDF1 was correlated with the TNM stage and T stage of patients with OSCC (P＜0. 05) ，and the patients with high expression of YTHDF1 had a shorter sur- vival time compared with those with low expression (P ＜0. 001) ．@*Conclusion @# High expression of YTHDF1 may be associated with poor patient prognosis and YTHDF1 may be able to serve as a target for OSCC treatment.
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Objective @#To investigate the value of methyltransferase-like protein 16 (METTL16) in the clinical di- agnosis and prognostic prediction of multiple myeloma (MM) patients . @*Methods @#The expression level and prog- nostic potential of each gene involved in N6 -methyladenosine ( m6A) modification in MM were respectively ana- lyzed in the databases of the Multiple Myeloma Research Foundation (MMRF) and the Genotype-Tissue Expression Proj ect (GTEx ) . Bone marrow specimens from 26 patients with initial diagnosis of MM and 19 patients with MM af- ter treatment with standard regimens and peripheral blood specimens from 24 normal subjects were collected respec- tively , and the expression levels of m6A genes were determined by qRT-PCR. The correlation between METTL16 expression and various laboratory and clinical indexes was analyzed: hemoglobin ( Hb) , white blood cell count ( WBC) , platelet count (PLT) , blood creatinine (Scr ) , serum calcium (Ca2 + ) , β-microglobulin ( β-MG) , bone destruction , ISS stage , type , and overall survival (OS) in the patients with primary diagnosis . The expression lev- els of interleukin (IL) -4 , IL-6 , IL-10 , IL-18 and chemokine ligand 2 ( CCL2) , CCL3 , CCL4 in the specimens were further examined and their correlation with the expression of METTL16 was investigated . @*Results @#Database analysis suggested that METTL16 expression was significantly higher in MM patient samples compared with normal controls , which was associated with poor prognosis and had certain diagnostic value . qRT-PCR results showed that the expression level of METTL16 in the bone marrow of patients with initial diagnosis of MM was significantly higher than that of treated patients and normal controls . Its expression was positively correlated with hemoglobin , leuko- cytes and stage , and its expression was positively correlated with CCL4 expression .@*Conclusion @#METTL16 expres- sion was significantly elevated in patients with MM , and its expression level was correlated with anemia , more bone destruction and worse stage , which might indicate a poor prognosis . The significant correlation between the expres- sion of METTL16 and CCL4 suggests that METTL16 may play a corresponding pathogenic role through the relevant pathway. METTL16 will have significant clinical value in the management of MM .
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N6-methyladenosine(m6A), the most common, abundant, and conserved RNA modification in eukaryotic cells, regulates RNA splicing, stability, output, degradation and translation through m6A methyltransferase, m6A demethylase, and m6A methylated binding proteins. Recent studies have found that abnormal m6A methylation may mediate a variety of pathological processes in eyes and participate in the occurrence and development of metabolic, inflammatory, degenerative ocular diseases and ocular tumors, such as diabetic retinopathy, cataract, age-related macular degeneration and uveal melanoma. This review aims to summarize the roles of m6A methylation modification in ocular cells and ocular diseases, elucidate the potential molecular mechanisms of m6A methylation in ocular diseases, so as to encourage innovative approaches in the treatment of these ocular diseases.
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Aim To investigate the role and potential mechanism of methyltransferase-like 5 (METTL5) in triple-negative breast cancer (TNBC) . Methods The expression of METTL5 in TNBC tumor tissues and cell lines was detected by immunohistochemistry and Western blot. After shRNA targeting METTL5 (shRNAMETTL5) was transfected into TNBC cells, cell proliferation, migration and invasion were detected by CCK-8, colony formation, wound healing and Transwell assays, respectively. Western blot was used to detect the expression of Wnt/p-catenin signaling-related key proteins. A xenograft tumor model was constructed to verify the effect of METTL5 knockdown on the growth of TNBC cells and Wnt/p-catenin signaling activity in vivo. Results The expression of METTL5 was up-regulated in TNBC tumor tissues and cell lines (P < 0. 01) . Knockdown of METTL5 significantly inhibited the proliferation, migration and invasion of TNBC cells and reduced the expression of Wnt/p-catenin signaling molecules (3-catenin, cyclin Dl, matrix metalloproteinase (MMP) -2 and MMP-7 (all P < 0. 01) . Knockdown of METTL5 reduced tumor growth and Wnt/pcatenin signaling activity in vivo. Conclusions Knockdown of METTL5 can inhibit the proliferation, migration and invasion of TNBC cells, which may be related to the inhibition of Wnt/p-catenin signaling pathway.
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Objective:To screen and verify the key radioresistance genes regulated by m6A methylation in nasopharyngeal carcinoma (NPC) based on the chip data and cell experiments.Methods:The microarray data of NPC radioresistance genes, m6A regulated genes and mRNA expression profiles of NPC genes were downloaded from Gene Expression Omnibus (GEO) database. The differential genes were screened and statistically analyzed by R software. The biological processes, signal pathways and interaction networks of these genes were analyzed by bioinformatics. The m6A regulatory factors were knocked down and the radioresistant strains were constructed. The above m6A differential radioresistant genes of NPC were screened and verified by real-time reverse transcription PCR (qRT-PCR) and Western blot. The m6A modification of screened genes and their direct binding ability with methyltransferase 3 (METTL3) were verified by methylated RNA immunoprecipitation qPCR (MeRIP-qPCR). The siRNA of selected genes was transfected into NPC cells, and after treatment with ionizing radiation, cell proliferation was detected by CCK-8 assay and EdU, apoptosis and cell cycle were detected by flow cytometry, and radiosensitivity was detected by clone formation assay. The trend of differences in the abundance of Fe 2+ and lipid peroxidation between the control and EGLN3 knockdown groups after ionizing radiation treatment was compared by paired t-test. Results:Chip data GSE48501 intersected with GSE200792 and GSE53819 to obtain 6 differential genes, including EGLN3, FOSL2, ADM, JUN, VEGFA and PRDM1. The target genes of EGLN3 and FOSL2 were further screened by TNMplot and KMplot, etc. The mRNA of the target genes directly bound to METTL3 and were subjected to its mediated modification of m6A. The target genes were up-regulated in the parental cells after irradiation in a dose and time gradient manner, which were also significantly up-regulated in radioresistant cells. After EGLN3 and FOSL2 were down regulated, the proliferation activity of NPC cells was more significantly decreased after irradiation, and the radiosensitization ratio was statistically significant compared with that of NPC cells without EGLN3 and FOSL2 down-regulation. After irradiation, EGLN3 down-regulated NPC cells significantly down-regulated glutathione peroxidase 4 (GPX4) expression, increased the abundance of Fe 2+ and lipid peroxidation, which played a role in radiosensitization by inducing ferroptosis. Conclusions:EGLN3 and FOSL2 play a role in radioresistance in NPC through METTL3 mediated m6A methylation. Down-regulation of EGLN3 combined with ionizing radiation can increase the intracellular Fe 2+ abundance and lipid peroxidation and down-reuglate the expression of GPX4 in NPC cells, which can enhance radiosensitization for NPC radiotherapy by the ferroptosis pathway.
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Objective @#To study the molecular mechanism of fat mass and obesity⁃associated protein (FTO) regulating hepatocellular carcinoma (HCC) . @*Methods@#HepG2 cells of knock⁃down FTO were constructed , HepG2 cells of knock⁃down FTO and HepG2 cells were collected , and high⁃throughput sequencing was performed using Illumina Hiseq platform to screen the gene expression differences between the two groups . Through GO and KEGG enrichment analysis of these differential genes , FTO regulatory pathways were studied and downstream target genes of FTO were screened . The role of FTO downstream target gene in HCC was revealed by bioinformatic analysis and cell experiments .@*Results@#Transcriptome sequencing showed that 386 genes were differentially expressed between HepG2 cells of knock⁃down FTO and HepG2 cells , and they were involved in biological processes such as response to interferon⁃gamma . The expression of IFIT2 , one of the most responsive interferon⁃stimulating genes , was up⁃regulated after FTO knockdown . Potential m6 A methylation occurred at multiple sites of IFIT2 . The survival of HCC patients with high expression of IFIT2 was significantly prolonged , and knock⁃down of IFIT2 promoted the growth and migration of HepG2 cells . @*Conclusion @#FTO may regulate IFIT2 by mediating m6 A , and further promote the occurrence and development of HCC .
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Objective@#To investigate GATA3 expression and the regulatory mechanism of m6A modification in the re- sponse of alveolar epithelial cells to radiation, and to provide a new therapeutic target for radiation-induced lung injury based on its pathogenesis.@*Methods@#Human lung epithelial cell line (A549) and mouse lung epithelial cell line (MLE-12) were exposed to X-ray irradiation with a single dose of 10 Gy (dose rate 1 Gy/min) and 6 Gy (dose rate 0.75 Gy/min), respect- ively. The expression of VIRMA gene (RNA methylase) was inhibited by lipofection of A549 cells and MLE-12 cells with shRNA-VIRMA plasmid and siRNA-VIRMA interfering fragment, respectively. Quantification of m6A RNA methylation was performed by colorimetry. Changes in the expression of mRNAs of VIRMA, GATA3, and epithelial-mesenchymal transition (EMT) markers in irradiated A549 and MLE-12 cells were determined by qRT-PCR. Changes in the expression of VIRMA, GATA3, and EMT marker proteins in irradiated A549 and MLE-12 cells were determined by Western blot.@*Results@#Radiation up-regulated the expression of methylase VIRMA in A549 and MLE-12 cells, which in turn enhanced the m6A of total RNA and the expression of GATA3 gene and protein, resulting in EMT. Furthermore, in A549 and MLE-12 cells, interference of the VIRMA gene significantly reduced the expression of GATA3 gene and protein and the expression of EMT-related molecules.@*Conclusion @#Radiation induces m6A modification in alveolar epithelial cells, which up-regu- lates the expression of GATA3 gene and induces EMT, thus playing an important role in the process of radiation-induced lung injury.
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AIM: To investigate the role and mechanism of N6-methyladenosine(m6A)methyltransferase 3(METTL3)in the pathogenesis of diabetic cataract.METHODS: We cultured SRA01/04 cells in low and high sugar media for 24h and measured changes in epithelial-mesenchymal transition(EMT)indicators(E-Cadherin, N-Cadherin, ZO-1 and α-SMA)using RT-qPCR and Western blot assays. Cell migration was also assessed using transwell and scratch assays. To investigate the expression level and localization of METTL3 in human lens anterior capsules tissues. Additionally, we used m6A dot blot assay to detect the m6A methylation level of cells cultured in low and high glucose media for 24h, and employed RT-qPCR and Western blot experiments to detect RNA and protein expression of METTL3 in cells. We then treated the cells with METTL3 inhibitor and measured changes in EMT markers by RT-qPCR and Western blot; m6A methylation level was detected by m6A dot blot test; cell migration was detected by Transwell. Finally, the expression of transforming growth factor-β(TGFβ1)in cultured cells was assessed by immunofluorescence staining and the expression levels of TGFβ1 and SNAIL in cells were determined using RT-qPCR and Western blot.RESULTS: Under high glucose conditions, the expression of EMT markers, METTL3, and m6A methylation levels were significantly increased in cells(P<0.05). Furthermore, the migratory ability of cells was higher in high-sugar medium than in low-sugar medium. In human lens anterior capsules, METTL3 expression was higher in patients with diabetic cataract compared to those with age-related cataract. Importantly, treatment with the METTL3 inhibitor STM2457 inhibited EMT in cells, the expression of TGFβ1 and SNAIL, as well as m6A methylation levels in cells(all P<0.05)compared to high-sugar + dimethyl sulfoxide(DMSO)group. Moreover, the migratory capacity of cells was reduced after the addition of STM2457 compared to the high-sugar + DMSO group.CONCLUSION:METTL3 promotes the EMT in human lens epithelial cells under high glucose conditions by activating the TGFβ1/SNAIL pathway, thus contributing to the development of diabetic cataracts.
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There are a variety of post-transcriptional modifications in mRNA, which regulate the stability, splicing, translation, transport and other processes of mRNA, followed by affecting cell development, body immunity, learning and cognition and other important physiological functions. m6A modification is one of the most abundant post-transcriptional modifications widely existing in mRNA, regulating the metabolic activities of RNA and affecting gene expression. m6A modified homeostasis is critical for the development and maintenance of the nervous system. In recent years, m6A modification has been found in neurodegenerative diseases, mental diseases and brain tumors. This review summarizes the role of m6A methylation modification in the development, function and related diseases of the central nervous system in recent years, providing potential clinical therapeutic targets for neurological diseases.
Subject(s)
Methylation , Central Nervous System/metabolism , RNA, Messenger/metabolism , RNAABSTRACT
The occurrence of gastric cancer is closely related to environmental, genetic, and epigenetic factors. Currently, RNA modification is a research frontier and hotspot in the field of epigenetics. With the advancements in analytical chemistry and high-throughput sequencing technologies, new technologies and methods of exploring RNA modification are constantly being presented. Numerous studies have confirmed the involvement of RNA modifications in the occurrence and development of various diseases. Recent studies have shown that RNA modifications such as m6A, m5C, and ac4C regulate the malignant progression of various tumors, including gastric cancer, liver cancer, colorectal cancer, and leukemia. This article systematically reviews the research status and mechanism of different RNA modifications in the occurrence and development of gastric cancer, as well as discusses its potential value in the diagnosis and treatment of gastric cancer.