{"title":"METTL3通过调节转录因子BACH2介导CPB1表达，促进晶状体上皮细胞凋亡和氧化应激。","authors":"Zhangxing Sheng, Yu Pan, Liqin Shao, Yihui Bao","doi":"10.1007/s10863-025-10054-1","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Cataracts are a significant cause of vision loss, adversely affecting the quality of human life. Numerous studies have reported that lens epithelial cells (LECs) play a crucial role in age-related cataract (ARC). However, the roles of carboxypeptidase B 1 (CPB1) and transcription factor BTB and CNC homologue 2 (BACH2) in the pathogenesis of ARC remain unclear. In this study, we aim to explore the contributions of CPB1 and BACH2 to the development of ARC.</p><p><strong>Methods: </strong>The Gene Expression Omnibus (GEO) was utilized to screen for differentially expressed genes. mRNA and protein levels were assessed using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis. Flow cytometry was conducted to analyze apoptosis. The levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and malondialdehyde (MDA) were measured using a commercial kit. Dual-luciferase reporter assays and chromatin immunoprecipitation (CHIP) were performed to investigate the interaction between CPB1 and BACH2. The methylation site of BACH2 was analyzed using the RNA-protein binding sites prediction suite and the sequence-based RNA adenosine methylation site predictor suite. Methylated RNA immunoprecipitation (Me-RIP) was employed to detect m6A modification level of BACH2.</p><p><strong>Results: </strong>In ARC and H<sub>2</sub>O<sub>2</sub>-induced human lens epithelial cells (HLECs), CPB1, BACH2, and METTL3 were found to be up-regulated. Silencing CPB1 reduced apoptosis and MDA levels while enhancing the activities of SOD and GSH-PX in H<sub>2</sub>O<sub>2</sub>-induced HLECs. Additionally, CPB1 was shown to bind to BACH2, and knockdown of BACH2 attenuated apoptosis and oxidative stress in H<sub>2</sub>O<sub>2</sub>-induced HLECs by targeting CPB1. Notably, METTL3 promoted the BACH2 expression by enhancing CPB1 expression in H<sub>2</sub>O<sub>2</sub>-induced HLECs. Finally, silencing METTL3 inhibited apoptosis and oxidative stress in H<sub>2</sub>O<sub>2</sub>-induced HLECs by hampering BACH2 expression.</p><p><strong>Conclusions: </strong>METTL3 facilitates apoptosis and oxidative stress in H<sub>2</sub>O<sub>2</sub>-induced HLECs by promoting the modification of BACH2 and CPB1 expression.</p>","PeriodicalId":15080,"journal":{"name":"Journal of Bioenergetics and Biomembranes","volume":" ","pages":""},"PeriodicalIF":2.9000,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"METTL3 mediates CPB1 expression by regulating transcription factor BACH2 to promote apoptosis and oxidative stress of lens epithelial cells.\",\"authors\":\"Zhangxing Sheng, Yu Pan, Liqin Shao, Yihui Bao\",\"doi\":\"10.1007/s10863-025-10054-1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Cataracts are a significant cause of vision loss, adversely affecting the quality of human life. Numerous studies have reported that lens epithelial cells (LECs) play a crucial role in age-related cataract (ARC). However, the roles of carboxypeptidase B 1 (CPB1) and transcription factor BTB and CNC homologue 2 (BACH2) in the pathogenesis of ARC remain unclear. In this study, we aim to explore the contributions of CPB1 and BACH2 to the development of ARC.</p><p><strong>Methods: </strong>The Gene Expression Omnibus (GEO) was utilized to screen for differentially expressed genes. mRNA and protein levels were assessed using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis. Flow cytometry was conducted to analyze apoptosis. The levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and malondialdehyde (MDA) were measured using a commercial kit. Dual-luciferase reporter assays and chromatin immunoprecipitation (CHIP) were performed to investigate the interaction between CPB1 and BACH2. The methylation site of BACH2 was analyzed using the RNA-protein binding sites prediction suite and the sequence-based RNA adenosine methylation site predictor suite. Methylated RNA immunoprecipitation (Me-RIP) was employed to detect m6A modification level of BACH2.</p><p><strong>Results: </strong>In ARC and H<sub>2</sub>O<sub>2</sub>-induced human lens epithelial cells (HLECs), CPB1, BACH2, and METTL3 were found to be up-regulated. Silencing CPB1 reduced apoptosis and MDA levels while enhancing the activities of SOD and GSH-PX in H<sub>2</sub>O<sub>2</sub>-induced HLECs. Additionally, CPB1 was shown to bind to BACH2, and knockdown of BACH2 attenuated apoptosis and oxidative stress in H<sub>2</sub>O<sub>2</sub>-induced HLECs by targeting CPB1. Notably, METTL3 promoted the BACH2 expression by enhancing CPB1 expression in H<sub>2</sub>O<sub>2</sub>-induced HLECs. Finally, silencing METTL3 inhibited apoptosis and oxidative stress in H<sub>2</sub>O<sub>2</sub>-induced HLECs by hampering BACH2 expression.</p><p><strong>Conclusions: </strong>METTL3 facilitates apoptosis and oxidative stress in H<sub>2</sub>O<sub>2</sub>-induced HLECs by promoting the modification of BACH2 and CPB1 expression.</p>\",\"PeriodicalId\":15080,\"journal\":{\"name\":\"Journal of Bioenergetics and Biomembranes\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":2.9000,\"publicationDate\":\"2025-02-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Bioenergetics and Biomembranes\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1007/s10863-025-10054-1\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOPHYSICS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Bioenergetics and Biomembranes","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s10863-025-10054-1","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOPHYSICS","Score":null,"Total":0}
METTL3 mediates CPB1 expression by regulating transcription factor BACH2 to promote apoptosis and oxidative stress of lens epithelial cells.
Background: Cataracts are a significant cause of vision loss, adversely affecting the quality of human life. Numerous studies have reported that lens epithelial cells (LECs) play a crucial role in age-related cataract (ARC). However, the roles of carboxypeptidase B 1 (CPB1) and transcription factor BTB and CNC homologue 2 (BACH2) in the pathogenesis of ARC remain unclear. In this study, we aim to explore the contributions of CPB1 and BACH2 to the development of ARC.
Methods: The Gene Expression Omnibus (GEO) was utilized to screen for differentially expressed genes. mRNA and protein levels were assessed using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis. Flow cytometry was conducted to analyze apoptosis. The levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and malondialdehyde (MDA) were measured using a commercial kit. Dual-luciferase reporter assays and chromatin immunoprecipitation (CHIP) were performed to investigate the interaction between CPB1 and BACH2. The methylation site of BACH2 was analyzed using the RNA-protein binding sites prediction suite and the sequence-based RNA adenosine methylation site predictor suite. Methylated RNA immunoprecipitation (Me-RIP) was employed to detect m6A modification level of BACH2.
Results: In ARC and H2O2-induced human lens epithelial cells (HLECs), CPB1, BACH2, and METTL3 were found to be up-regulated. Silencing CPB1 reduced apoptosis and MDA levels while enhancing the activities of SOD and GSH-PX in H2O2-induced HLECs. Additionally, CPB1 was shown to bind to BACH2, and knockdown of BACH2 attenuated apoptosis and oxidative stress in H2O2-induced HLECs by targeting CPB1. Notably, METTL3 promoted the BACH2 expression by enhancing CPB1 expression in H2O2-induced HLECs. Finally, silencing METTL3 inhibited apoptosis and oxidative stress in H2O2-induced HLECs by hampering BACH2 expression.
Conclusions: METTL3 facilitates apoptosis and oxidative stress in H2O2-induced HLECs by promoting the modification of BACH2 and CPB1 expression.
期刊介绍：
The Journal of Bioenergetics and Biomembranes is an international journal devoted to the publication of original research that contributes to fundamental knowledge in the areas of bioenergetics, biomembranes, and transport, including oxidative phosphorylation, photosynthesis, muscle contraction, as well as cellular and systemic metabolism. The timely research in this international journal benefits biophysicists, membrane biologists, cell biologists, biochemists, molecular biologists, physiologists, endocrinologists, and bio-organic chemists.
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