Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis最新文献

Deciphering the structural and functional impact of interleukin-3 (IL-3) variants: Insights from site-directed mutagenesis, computational modeling, molecular dynamics simulations, and implications for anticancer therapies and immune regulation 解读白介素-3 （IL-3）变异的结构和功能影响：来自定点突变、计算建模、分子动力学模拟的见解，以及抗癌治疗和免疫调节的意义。

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-12-13 DOI: 10.1016/j.mrfmmm.2025.111927

Allah Rakha Yaseen , Uzma Nisar , Munir Ahmad , Faiza Saleem , Hafsa Amjad

Interleukin-3 (IL-3) is a pivotal cytokine in immune regulation, making it a compelling target for therapeutic intervention in immune-related disorders. This study employs an integrative computational approach, combining site-directed mutagenesis, molecular dynamics simulations, and protein-protein docking, to systematically investigate the structural and functional consequences of IL-3 mutations. By mapping disease-associated variants and their mechanistic impact on IL-3 receptor (IL-3Rα) interactions, we uncover critical determinants of cytokine signaling and stability. Our findings reveal that mutations such as E41L, R127L, and W123E induce structural perturbations linked to disease phenotypes, underscoring their potential as therapeutic targets. Conversely, variants like H45R, D40A, and N124D exhibit altered receptor-binding affinities, presenting opportunities for rational drug design. Protein-protein docking further elucidates the molecular basis of IL-3Rα recognition, highlighting key residues that govern binding specificity. Notably, mutations affecting interface residues drive substantial changes in binding energetics, providing novel insights into IL-3-mediated signaling modulation. This study advances the understanding of IL-3 structural dynamics through a computational lens, offering a predictive framework for mutation-driven functional alterations. By integrating biophysical modeling with functional annotation, we establish a foundation for precision engineering of IL-3 variants with therapeutic potential. Despite the inherent limitations of in silico modeling, our approach provides preliminary computational insights into cytokine modulation and forms a foundation for future experimental validation aimed at exploring potential therapeutic avenues for IL-3-associated pathologies

白细胞介素-3 （IL-3）是免疫调节中的关键细胞因子，使其成为免疫相关疾病治疗干预的一个引人注目的靶点。本研究采用综合计算方法，结合位点定向诱变、分子动力学模拟和蛋白-蛋白对接，系统研究IL-3突变的结构和功能后果。通过绘制疾病相关变异及其对IL-3受体（IL-3Rα）相互作用的机制影响，我们揭示了细胞因子信号传导和稳定性的关键决定因素。我们的研究结果表明，突变如E41L、R127L和W123E诱导与疾病表型相关的结构扰动，强调了它们作为治疗靶点的潜力。相反，H45R、D40A和N124D等变体表现出改变的受体结合亲和力，为合理的药物设计提供了机会。蛋白-蛋白对接进一步阐明了IL-3Rα识别的分子基础，突出了控制结合特异性的关键残基。值得注意的是，影响界面残基的突变驱动结合能量学的实质性变化，为il -3介导的信号调节提供了新的见解。本研究通过计算透镜推进了对IL-3结构动力学的理解，为突变驱动的功能改变提供了预测框架。通过将生物物理建模与功能注释相结合，我们为具有治疗潜力的IL-3变异的精确工程奠定了基础。尽管计算机模拟的固有局限性，我们的方法为细胞因子调节提供了初步的计算见解，并为未来的实验验证奠定了基础，旨在探索il -3相关病理的潜在治疗途径。

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引用次数: 0

SPDEF-mediated BIRC5 transcriptional activation enhances non-small cell lung cancer progression spdef介导的BIRC5转录激活促进非小细胞肺癌的进展

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-12-12 DOI: 10.1016/j.mrfmmm.2025.111926

Zhixiong Qiao , Zhiping Lv , Xuyang Song, Jinxi He

Background

SAM-pointed domain-containing ETS transcription factor (SPDEF), an ETS family transcription factor (TF), functions as a tumor-promoting factor in non-small cell lung cancer (NSCLC). However, the precise mechanisms of SPDEF in regulating NSCLC pathogenesis remain to be fully characterized.

Methods

Bioinformatics analyses were used to identify disease-associated TFs and downstream targets. A protein-protein interaction (PPI) network was constructed to predict hub TFs. The relationship between SPDEF and BIRC5 was validated by luciferase and ChIP assays. The influences on cell phenotypes were evaluated by testing cell colony formation, invasion, migration, and apoptosis. Xenograft studies were used to explore the role in tumor growth.

Results

This study revealed 101 disease-associated TFs in NSCLC. GO enrichment analysis of these TFs demonstrated significant enrichment in biological processes including lung development, lung differentiation, and Wnt signaling pathway. PPI analysis revealed FOXA1, GATA6, TBX2, and SPDEF as core regulatory factors. SPDEF and BIRC5 levels were upregulated in NSCLC tumors and cell lines. Mechanistically, SPDEF induced BIRC5 upregulation through transcriptional regulation. SPDEF depletion impaired NSCLC cell invasion, migration, and clonogenicity while inducing apoptosis in vitro, which could be abolished by BIRC5 reconstitution. Moreover, SPDEF deficiency inhibited A549 tumor growth in subcutaneous xenograft models. Additionally, BIRC5 activated the Wnt/β-catenin pathway in NSCLC cells.

Conclusion

Our findings demonstrate the tumor-promoting function of SPDEF in NSCLC by transcriptionally activating BIRC5, highlighting SPDEF as a candidate target for future NSCLC-directed therapies.

背景：含sam点结构域的ETS转录因子（SPDEF）是ETS家族转录因子（TF）的一种，在非小细胞肺癌（NSCLC）中发挥肿瘤促进因子的作用。然而，SPDEF调控NSCLC发病机制的确切机制尚不清楚。方法采用生物信息学方法鉴定与疾病相关的tf和下游靶点。构建了蛋白相互作用（PPI）网络来预测轮毂TFs。SPDEF与BIRC5之间的关系通过荧光素酶和ChIP检测得到验证。通过检测细胞集落形成、侵袭、迁移和凋亡来评估对细胞表型的影响。异种移植研究用于探索其在肿瘤生长中的作用。结果本研究发现101例非小细胞肺癌相关肿瘤转移灶。氧化石墨烯富集分析表明，这些tf在肺发育、肺分化和Wnt信号通路等生物过程中显著富集。PPI分析显示FOXA1、GATA6、TBX2和SPDEF是核心调控因子。SPDEF和BIRC5水平在NSCLC肿瘤和细胞系中上调。机制上，SPDEF通过转录调控诱导BIRC5上调。SPDEF缺失在体外诱导细胞凋亡的同时，损害了NSCLC细胞的侵袭、迁移和克隆原性，而这一过程可以通过BIRC5重构来消除。此外，SPDEF缺乏抑制皮下异种移植模型中A549肿瘤的生长。此外，BIRC5在NSCLC细胞中激活Wnt/β-catenin通路。我们的研究结果表明SPDEF通过转录激活BIRC5在NSCLC中具有促肿瘤功能，突出了SPDEF作为未来NSCLC定向治疗的候选靶点。

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引用次数: 0

Computational analysis of functional, structural and pathogenic impacts of missense SNPs in the human SOCS3 gene 人类SOCS3基因错义snp对功能、结构和致病影响的计算分析。

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-12-11 DOI: 10.1016/j.mrfmmm.2025.111925

Raviteja Reddy Alipeddi, Durga Neeharika Rani, Pallavi Sampanmudumby, Surekha Rani Hanumanth

The Suppressor of Cytokine Signaling 3 (SOCS3) protein is a crucial negative regulator of cytokine-mediated signaling pathways, and genetic variations in SOCS3 have been implicated in the development of multiple multi-factorial diseases (MFDs). In this study, a comprehensive insilico analysis was performed to evaluate the functional and structural consequences of eight missense single nucleotide polymorphisms (SNPs) (rs37610117, rs111889212, rs1061489, rs376015024, rs17849241, rs201763454, rs200504273, rs150709546) retrieved from the NCBI dbSNP database. Functional predictions using SIFT, PolyPhen-2, PhD-SNP, PANTHER, and SNP&GO identified several potentially deleterious variants, with rs201763454 (L156F) notably predicted to impair protein function. Conservation analysis with ConSurf revealed that key residues such as H126Y and A223S are highly conserved, indicating their functional importance. Homology-based structural modeling via Phyre2 indicated SNP-induced alterations in secondary structure, while stability and flexibility assessments suggested destabilizing effects for multiple variants. Structural mapping confirmed that high-risk SNPs are localized within critical functional domains of SOCS3. CScape cancer susceptibility prediction revealed that E98V and A223S as oncogenic SNPs. Further analysis using cBioPortal and CanSAR Black confirmed the involvement of E98V and other variants (S26N, F136L, A223S) in various cancers, highlighting their potential clinical significance. Protein-protein interaction analyses (STRING, GeneMANIA) further highlighted the biological relevance. Collectively, this study identifies critical SOCS3 SNPs that may modulate protein function and contribute to MFD pathogenesis. These findings suggest that SOCS3 missense SNPs, particularly E98V, H126Y, and A223S, may serve as candidate biomarkers for cancer susceptibility and warrant further experimental validation

细胞因子信号传导抑制因子3 （SOCS3）蛋白是细胞因子介导的信号传导途径的关键负调控因子，SOCS3的遗传变异与多种多因子疾病（mfd）的发展有关。在这项研究中，对从NCBI dbSNP数据库中检索到的8个错义单核苷酸多态性（rs37610117、rs111889212、rs1061489、rs376015024、rs17849241、rs201763454、rs200504273、rs150709546）的功能和结构后果进行了全面的计算机分析。使用SIFT、polyphen2、PhD-SNP、PANTHER和SNP&GO进行功能预测，发现了几个潜在的有害变异，其中rs201763454 （L156F）被预测会损害蛋白质功能。ConSurf保守分析显示，H126Y和A223S等关键残基高度保守，表明其功能重要性。通过Phyre2进行的基于同源性的结构建模表明snp诱导的二级结构改变，而稳定性和灵活性评估表明多种变异的不稳定效应。结构图谱证实，高风险snp位于SOCS3的关键功能域内。CScape癌症易感性预测显示E98V和A223S为致癌snp。使用cbiopportal和canansar Black进一步分析证实了E98V和其他变体（S26N、F136L、A223S）参与多种癌症，强调了它们潜在的临床意义。蛋白-蛋白相互作用分析（STRING, GeneMANIA）进一步强调了生物学相关性。总的来说，本研究确定了可能调节蛋白质功能并促进MFD发病机制的关键SOCS3 snp。这些发现表明，SOCS3错义snp，特别是E98V、H126Y和A223S，可能作为癌症易感性的候选生物标志物，需要进一步的实验验证。

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引用次数: 0

Role of c-Myc-regulated miR-196a-5p in the progression of triple-negative breast cancer: Potential involvement of HOXA7 and HOXB7 c- myc调控的miR-196a-5p在三阴性乳腺癌进展中的作用：HOXA7和HOXB7的潜在参与

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-12-05 DOI: 10.1016/j.mrfmmm.2025.111923

Xin-Yu Song , Le Yang , Hui-Ling Li , Hao-Yi Xu , Xiao-Li Xu , Kang Yan , Yuan-Jing Liu , Zuliyaer Mierzhakenmu , Rui Xu , Chao Dong

Objective

This study aimed to elucidate the regulatory role of c-Myc-associated miR-196a-5p in the progression of triple-negative breast cancer (TNBC). We further explored its target genes to investigate possible mechanisms of action.

Methods

High-throughput sequencing was employed to profile microRNA expression in MDA-MB-231 cells with differential c-Myc expression. Differentially expressed microRNAs were identified, and miR-196a-5p levels, along with the expression of predicted target genes, were quantified using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in c-Myc knockdown cells. Candidate targets were predicted using miRDB and TargetScan databases and further validated by qRT-PCR and Western blotting in MDA-MB-231 and MDA-MB-468 cells with modulated miR-196a-5p expression. Functional assays, including clonogenic assays, flow cytometry, and Transwell migration and invasion assays were conducted to evaluate the effects of miR-196a-5p on cellular behavior.

Results

miR-196a-5p expression was significantly reduced in c-Myc-deficient cells and exhibited a positive association with c-Myc levels. Bioinformatic analysis identified HOXA7 and HOXB7 as miR-196a-5p putative targets, with inverse expression patterns observed between the microRNA and these genes. Downregulation of miR-196a-5p resulted in elevated expression of HOXA7 and HOXB7, whereas its overexpression resulted in their suppression. Functionally, increased levels of miR-196a-5p were associated with enhanced proliferation, invasion, and migration, along with decreased apoptosis in TNBC cells.

Conclusion

Our findings reveal a positive correlation between c-Myc and miR-196a-5p in TNBC, suggesting that miR-196a-5p may promote tumor progression by regulating cellular proliferation, invasion, migration, and apoptosis. The inverse expression patterns between miR-196a-5p and HOXA7/HOXB7 imply these genes may be downstream targets, though direct causal relationships and detailed molecular mechanisms require further validation.

目的本研究旨在阐明c- myc相关的miR-196a-5p在三阴性乳腺癌（TNBC）进展中的调节作用。我们进一步探索其靶基因，探讨其可能的作用机制。方法采用高通量测序方法对c-Myc表达差异的MDA-MB-231细胞进行microRNA表达分析。鉴定差异表达的microrna，并在c-Myc敲低细胞中使用定量逆转录聚合酶链反应（qRT-PCR）定量miR-196a-5p水平以及预测靶基因的表达。使用miRDB和TargetScan数据库预测候选靶点，并在miR-196a-5p表达调节的MDA-MB-231和MDA-MB-468细胞中通过qRT-PCR和Western blotting进一步验证。通过功能分析，包括克隆实验、流式细胞术、Transwell迁移和侵袭实验来评估miR-196a-5p对细胞行为的影响。结果smir -196a-5p在c-Myc缺陷细胞中表达显著降低，并与c-Myc水平呈正相关。生物信息学分析发现HOXA7和HOXB7是miR-196a-5p的推测靶点，在microRNA和这些基因之间观察到相反的表达模式。miR-196a-5p的下调导致HOXA7和HOXB7的表达升高，而miR-196a-5p的过表达导致HOXA7和HOXB7的抑制。在功能上，miR-196a-5p水平的升高与TNBC细胞的增殖、侵袭和迁移增强以及凋亡减少有关。我们的研究结果显示，TNBC中c-Myc与miR-196a-5p呈正相关，表明miR-196a-5p可能通过调节细胞增殖、侵袭、迁移和凋亡来促进肿瘤进展。miR-196a-5p与HOXA7/HOXB7之间的反向表达模式暗示这些基因可能是下游靶点，但直接因果关系和详细的分子机制需要进一步验证。

{"title":"Role of c-Myc-regulated miR-196a-5p in the progression of triple-negative breast cancer: Potential involvement of HOXA7 and HOXB7","authors":"Xin-Yu Song , Le Yang , Hui-Ling Li , Hao-Yi Xu , Xiao-Li Xu , Kang Yan , Yuan-Jing Liu , Zuliyaer Mierzhakenmu , Rui Xu , Chao Dong","doi":"10.1016/j.mrfmmm.2025.111923","DOIUrl":"10.1016/j.mrfmmm.2025.111923","url":null,"abstract":"<div><h3>Objective</h3><div>This study aimed to elucidate the regulatory role of c-Myc-associated miR-196a-5p in the progression of triple-negative breast cancer (TNBC). We further explored its target genes to investigate possible mechanisms of action.</div></div><div><h3>Methods</h3><div>High-throughput sequencing was employed to profile microRNA expression in MDA-MB-231 cells with differential c-Myc expression. Differentially expressed microRNAs were identified, and miR-196a-5p levels, along with the expression of predicted target genes, were quantified using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in c-Myc knockdown cells. Candidate targets were predicted using miRDB and TargetScan databases and further validated by qRT-PCR and Western blotting in MDA-MB-231 and MDA-MB-468 cells with modulated miR-196a-5p expression. Functional assays, including clonogenic assays, flow cytometry, and Transwell migration and invasion assays were conducted to evaluate the effects of miR-196a-5p on cellular behavior.</div></div><div><h3>Results</h3><div>miR-196a-5p expression was significantly reduced in c-Myc-deficient cells and exhibited a positive association with c-Myc levels. Bioinformatic analysis identified HOXA7 and HOXB7 as miR-196a-5p putative targets, with inverse expression patterns observed between the microRNA and these genes. Downregulation of miR-196a-5p resulted in elevated expression of HOXA7 and HOXB7, whereas its overexpression resulted in their suppression. Functionally, increased levels of miR-196a-5p were associated with enhanced proliferation, invasion, and migration, along with decreased apoptosis in TNBC cells.</div></div><div><h3>Conclusion</h3><div>Our findings reveal a positive correlation between c-Myc and miR-196a-5p in TNBC, suggesting that miR-196a-5p may promote tumor progression by regulating cellular proliferation, invasion, migration, and apoptosis. The inverse expression patterns between miR-196a-5p and HOXA7/HOXB7 imply these genes may be downstream targets, though direct causal relationships and detailed molecular mechanisms require further validation.</div></div>","PeriodicalId":49790,"journal":{"name":"Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis","volume":"832 ","pages":"Article 111923"},"PeriodicalIF":1.9,"publicationDate":"2025-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145738425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

DNA double-strand breaks induced by DNA topoisomerase IIα and reactive oxygen species lead to the integration of foreign DNA into the host genome DNA拓扑异构酶i α和活性氧诱导的DNA双链断裂导致外源DNA整合到宿主基因组中

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-12-04 DOI: 10.1016/j.mrfmmm.2025.111924

Aya Kurosawa , Masumi Umehara , Noritaka Adachi

Foreign DNA integrates into the genome at low frequencies and random positions upon introduction into cells. This phenomenon, known as random integration, occurs when DNA double-strand breaks (DSBs) are repaired by non-homologous end-joining or polymerase theta-mediated end-joining, incorporating the foreign DNA into the genome. However, the mechanism underlying DSB generation in random integration remains unclear. In this study, we investigated the role of DNA topoisomerase II (Top2) and reactive oxygen species (ROS) in generating DSBs and examined their effects on random integration frequency. In Nalm-6 cells, the frequency of random integration increased following treatment with the Top2 inhibitor etoposide and hydrogen peroxide. Conversely, depletion of Top2α and cultivation under hypoxic conditions independently reduced the frequency of random integration, with their combination resulting in an even greater reduction. In gene targeting experiments at the HPRT locus, both Top2α depletion and hypoxic culture similarly reduced random integration. This suggests that DSBs generated by Top2α and ROS contribute to random integration and impede efficient gene targeting.

外源DNA在进入细胞后以低频率和随机位置整合到基因组中。这种被称为随机整合的现象发生在DNA双链断裂（dsb）通过非同源末端连接或聚合酶介导的末端连接修复时，将外源DNA整合到基因组中。然而，DSB在随机整合中产生的机制尚不清楚。在这项研究中，我们研究了DNA拓扑异构酶II （Top2）和活性氧（ROS）在dsb生成中的作用，并研究了它们对随机整合频率的影响。在Nalm-6细胞中，用Top2抑制剂依托泊苷和过氧化氢处理后，随机整合的频率增加。相反，Top2α的消耗和缺氧条件下的培养单独降低了随机整合的频率，它们的结合导致更大的减少。在HPRT基因位点的基因靶向实验中，Top2α缺失和缺氧培养同样减少了随机整合。这表明由Top2α和ROS产生的dsb促进了随机整合，阻碍了有效的基因靶向。

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引用次数: 0

Deciphering the molecular mechanism of YY1/HIF1A modulating ovarian cancer angiogenesis based on single-cell transcriptomics technology 基于单细胞转录组学技术解读YY1/HIF1A调控卵巢癌血管生成的分子机制

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-07-01 DOI: 10.1016/j.mrfmmm.2025.111916

Xiyun Quan , Huimei Yi , Meiyuan Huang, Dongliang Chen

Background

Angiogenesis assumes an essential role in tumor development and is a fundamental condition for tumor growth. Yin Yang 1 (YY1) is highly expressed in various types of cancers and is a key player in tumor angiogenesis, but its role in ovarian cancer (OC) has not been fully elucidated. Therefore, this study will delve into the mechanism of YY1 in OC angiogenesis.

Methods

Based on single-cell transcriptomics data of OC tumor samples and adjacent samples downloaded from the GEO database, differentially expressed genes (DEGs) and related signaling pathways were screened and validated in OC cells. Furthermore, co-culture technology was applied to assess the impact of YY1 expression in OC cells on angiogenesis ability. The molecular mechanism of YY1 regulation of OC angiogenesis was explored through bioinformatics analysis combined with co-immunoprecipitation, chromatin immunoprecipitation, and dual-luciferase reporter gene assays. Rescue experiments were designed, with results validated in qRT-PCR, angiogenesis assays, and Western blotting.

Results

Based on re-analysis of single-cell transcriptomics data from OC tumor samples and adjacent samples, we found that YY1 expression was significantly upregulated in OC cells, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis results showed that DEGs in YY1-positive tumor cells were significantly enriched in the HIF-1 signaling pathway. Moreover, in vitro experiments demonstrated that YY1 was highly expressed in OC to boost OC angiogenesis. Specifically, YY1 can stabilize hypoxia-inducible factor 1α (HIF1A) expression by competitively binding to WD repeat domain-containing 7 (FBXW7), thereby facilitating the transcriptional activation of angiogenesis genes. Finally, we demonstrated through rescue experiments that targeting the YY1/HIF1A axis can repress OC angiogenesis.

Conclusion

Through single-cell transcriptomics analysis combined with cell experiments, we proved the specific mechanism by which YY1 affects the angiogenesis ability of OC. YY1 affects the expression of angiogenesis genes by modulating the FBXW7/HIF1A axis.

肿瘤发生在肿瘤的发展过程中起着至关重要的作用，是肿瘤生长的基本条件。阴阳1 （YY1）在各种类型的癌症中高表达，是肿瘤血管生成的关键参与者，但其在卵巢癌（OC）中的作用尚未完全阐明。因此，本研究将深入探讨YY1在OC血管生成中的作用机制。方法基于从GEO数据库下载的OC肿瘤样本及邻近样本的单细胞转录组学数据，筛选OC细胞中的差异表达基因（differential expression genes, DEGs）及相关信号通路并进行验证。此外，采用共培养技术评估YY1在OC细胞中的表达对血管生成能力的影响。通过生物信息学分析，结合共免疫沉淀、染色质免疫沉淀、双荧光素酶报告基因检测，探讨YY1调控OC血管生成的分子机制。设计了救援实验，并通过qRT-PCR、血管生成实验和Western blotting验证了结果。结果通过对OC肿瘤样本和邻近样本的单细胞转录组学数据的重新分析，我们发现YY1在OC细胞中的表达显著上调，基因本体（GO）和京都基因基因组百科全书（KEGG）富集分析结果显示YY1阳性肿瘤细胞中的DEGs在HIF-1信号通路中显著富集。此外，体外实验表明，YY1在OC中高表达，促进OC血管生成。具体来说，YY1可以通过竞争性结合WD重复结构域7 （FBXW7）来稳定缺氧诱导因子1α (HIF1A)的表达，从而促进血管生成基因的转录激活。最后，我们通过救援实验证明，靶向YY1/HIF1A轴可以抑制OC血管生成。结论通过单细胞转录组学分析结合细胞实验，证实了YY1影响OC血管生成能力的具体机制。YY1通过调节FBXW7/HIF1A轴影响血管生成基因的表达。

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引用次数: 0

FOXA2 promotes glutamine metabolism to facilitate the malignant development of bladder cancer by transcriptionally increasing GLS1 expression FOXA2通过转录增加GLS1的表达，促进谷氨酰胺代谢，促进膀胱癌的恶性发展。

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-07-01 DOI: 10.1016/j.mrfmmm.2025.111920

Quan Yuan , Bowen Liu , Wei Yan , Huifeng Li , Jinxian Pu

Background

Forkhead box A2 (FOXA2) is found to be abnormally overexpressed in bladder cancer (BCa), but its role and underlying molecular mechanisms in BCa progression remain revealed.

Methods

The expression levels of FOXA2, glutaminase 1 (GLS1), glutamine metabolism-related markers were examined using qRT-PCR and western blot. Glutamine metabolism was assessed by detecting glutamine uptake, intracellular glutamate, ATP, GSH and ROS levels. BCa cell proliferation, migration and invasion were analyzed by CCK8 assay, EdU assay, wound healing assay, and Transwell assay. The regulation of FOXA2 on GLS1 promoter was confirmed by dual-luciferase reporter assay and ChIP assay. Mice xenograft tumor models were constructed to evaluate the role of FOXA2 in BCa tumorigenesis.

Results

FOXA2 expression was upregulated in BCa cells and its knockdown significantly decreased GLS1 expression. Silencing of FOXA2 inhibited BCa cell glutamine metabolism, thus suppressing cell proliferation and metastasis, and these effects were reversed by GLS1 overexpression. In terms of mechanism, FOXA2 increased the transcription and expression of GLS1 by binding to its promoter region. Animal study revealed that FOXA2 interference also reduced BCa tumorigenesis through decreasing GLS1 expression.

Conclusion

FOXA2 accelerated BCa cell proliferation and metastasis by promoting GLS1-mediated glutamine metabolism, providing a novel therapy target for BCa.

背景：叉头盒A2 （FOXA2）在膀胱癌（BCa）中被发现异常过表达，但其在BCa进展中的作用和潜在的分子机制尚不清楚。方法：采用qRT-PCR和western blot检测FOXA2、谷氨酰胺酶1 （GLS1）、谷氨酰胺代谢相关标志物的表达水平。通过检测谷氨酰胺摄取、细胞内谷氨酸、ATP、GSH和ROS水平来评估谷氨酰胺代谢。CCK8法、EdU法、创面愈合法和Transwell法分析BCa细胞的增殖、迁移和侵袭。双荧光素酶报告子实验和ChIP实验证实FOXA2对GLS1启动子的调控作用。构建小鼠异种移植瘤模型，评价FOXA2在BCa肿瘤发生中的作用。结果：FOXA2在BCa细胞中表达上调，敲低FOXA2可显著降低GLS1的表达。FOXA2的沉默抑制了BCa细胞谷氨酰胺代谢，从而抑制了细胞的增殖和转移，这些作用被GLS1过表达逆转。从机制上看，FOXA2通过结合GLS1的启动子区增加了GLS1的转录和表达。动物实验表明FOXA2的干扰也通过降低GLS1的表达来减少BCa的肿瘤发生。结论：FOXA2通过促进gls1介导的谷氨酰胺代谢，促进BCa细胞的增殖和转移，为BCa提供了新的治疗靶点。

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引用次数: 0

Implications of seven NTCP mutations for receptor stability and Hepatitis B Virus infectivity: A computational analysis 7种NTCP突变对受体稳定性和乙型肝炎病毒传染性的影响：计算分析。

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-07-01 DOI: 10.1016/j.mrfmmm.2025.111921

Amina Kardoudi , Salaheddine Redouane , Salma Madihi , Thomas Jackson , Abdelouaheb Benani

Hepatitis B is a widespread viral infection and a major global public health concern. The human sodium taurocholate co-transporter polypeptide (NTCP) serves as a key receptor for the hepatitis B virus (HBV), enabling its entry into hepatocytes. Understanding how specific NTCP mutations influence its stability and interaction with HBV is critical for elucidating mechanisms of viral infectivity and resistance. This study evaluates the impact of seven non-synonymous NTCP mutations on receptor stability and HBV binding using a comprehensive bioinformatics approach. Mutant NTCP/HBsAg complexes were generated via HADDOCK, and binding affinities were predicted using PRODIGY. Molecular dynamics simulations with GROMACS further assessed the stability and behavior of NTCP/PreS1 complexes. Our findings reveal that V160M and S267F significantly reduce complex stability and binding affinity, suggesting a potential role in natural resistance to HBV infection. Mutations I88T and R84W moderately affect NTCP–HBV interactions, while V200M, I223T, and I279T show minimal impact, maintaining wild-type reference complex characteristics. This study highlights the differential effects of NTCP mutations on HBV infectivity, providing insights into host susceptibility and resistance. The integrative computational strategy offers a robust framework for understanding HBV-host interactions and may aid in identifying novel therapeutic targets.

乙型肝炎是一种广泛的病毒感染，也是一个主要的全球公共卫生问题。人牛磺酸胆酸钠共转运蛋白多肽（NTCP）是乙型肝炎病毒（HBV）的关键受体，使其能够进入肝细胞。了解特异性NTCP突变如何影响其稳定性和与HBV的相互作用，对于阐明病毒感染和耐药机制至关重要。本研究利用综合生物信息学方法评估了七种非同义NTCP突变对受体稳定性和HBV结合的影响。通过HADDOCK产生突变的NTCP/HBsAg复合物，并用PRODIGY预测其结合亲和力。利用GROMACS进行分子动力学模拟，进一步评估了NTCP/PreS1复合物的稳定性和行为。我们的研究结果表明，V160M和S267F显著降低了复合物的稳定性和结合亲和力，提示在HBV感染的自然抗性中发挥潜在作用。突变I88T和R84W中度影响NTCP-HBV相互作用，而v2m、I223T和I279T影响最小，保持野生型参考复合物特征。这项研究强调了NTCP突变对HBV感染性的不同影响，为宿主易感性和耐药性提供了见解。综合计算策略为理解hbv -宿主相互作用提供了一个强大的框架，并可能有助于确定新的治疗靶点。

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引用次数: 0

WTAP-mediated m6A modification of ARG2 mRNA Inhibits Its expression and drives prostate cancer malignant progression wtap介导的m6A修饰ARG2 mRNA抑制其表达并驱动前列腺癌恶性进展

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-07-01 DOI: 10.1016/j.mrfmmm.2025.111912

Jianxin Li , Yu Zheng , Chaojiang Chen , Zhuoyuan Lin , Jiangang Pan , Funeng Jiang , Weide Zhong

Background

Prostate cancer (PCa) incidence increases as age advances and seriously endangers men’s health worldwide. Arginase 2 (ARG2) has been identified as a potential diagnostic and prognostic marker for PCa. However, the molecular mechanisms underlying its function in PCa remain undefined.

Methods

ARG2 mRNA and protein expression were quantified in PCa tissues and cells using qRT-PCR and Western blot. Cellular proliferation, glucose consumption, lactate production, apoptosis, and ferroptosis were evaluated via EdU incorporation, colony formation assays, commercial kits, and flow cytometry. Subsequently, the xenograft model was established to assess ARG2’s role in tumor growth in vivo. Bioinformatics analysis and RNA immunoprecipitation (RIP) were employed to investigate the interaction between Wilms’ tumor 1-associating protein (WTAP), a key component of the N6-methyladenosine (m6A) methyltransferase complex, and ARG2 mRNA. Besides, mRNA stability was determined using actinomycin D chase assays.

Results

ARG2 exhibited low expression in PCa tissues and cells. Upregulation of ARG2 inhibited proliferation and glycolysis, and promoted apoptosis, oxidative stress and ferroptosis of PCa cells. However, silencing ARG2 had the opposite effects. In vivo, ARG2 overexpression suppressed tumor growth. Mechanistically, WTAP bound directly to ARG2 mRNA, and their expression levels were inversely correlated. WTAP knockdown phenocopied ARG2 overexpression by repressing proliferation and glycolysis and enhancing apoptosis/ferroptosis, effects reversed by ARG2 silencing. ARG2 overexpression counteracted the oncogenic effects of WTAP overexpression.

Conclusion

WTAP bound to ARG2 and suppressed its expression, thereby promoting the malignant progression of PCa.

前列腺癌（PCa）的发病率随着年龄的增长而增加，严重危害着全球男性的健康。精氨酸酶2 （ARG2）已被确定为前列腺癌的潜在诊断和预后标志物。然而，其在前列腺癌中作用的分子机制仍不明确。方法采用qRT-PCR和Western blot方法检测前列腺癌组织和细胞中sarg2 mRNA和蛋白的表达。细胞增殖、葡萄糖消耗、乳酸生成、凋亡和铁下垂通过EdU并入、菌落形成测定、商业试剂盒和流式细胞术进行评估。随后，我们建立了异种移植物模型来评估ARG2在体内肿瘤生长中的作用。采用生物信息学分析和RNA免疫沉淀（RIP）技术研究了n6 -甲基腺苷（m6A）甲基转移酶复合物的关键组分Wilms ' tumor 1- associated protein （WTAP）与ARG2 mRNA的相互作用。此外，利用放线菌素D追逐法测定mRNA的稳定性。结果sarg2在前列腺癌组织和细胞中低表达。ARG2的上调抑制了PCa细胞的增殖和糖酵解，促进了细胞凋亡、氧化应激和铁下垂。然而，沉默ARG2具有相反的效果。在体内，ARG2过表达抑制肿瘤生长。在机制上，WTAP直接与arg2mrna结合，两者的表达水平呈负相关。WTAP通过抑制增殖和糖酵解以及增强细胞凋亡/铁下垂来抑制表型ARG2的过表达，而这种作用被ARG2沉默逆转。ARG2过表达抵消了WTAP过表达的致癌作用。结论wtap与ARG2结合，抑制其表达，从而促进前列腺癌的恶性进展。

{"title":"WTAP-mediated m6A modification of ARG2 mRNA Inhibits Its expression and drives prostate cancer malignant progression","authors":"Jianxin Li , Yu Zheng , Chaojiang Chen , Zhuoyuan Lin , Jiangang Pan , Funeng Jiang , Weide Zhong","doi":"10.1016/j.mrfmmm.2025.111912","DOIUrl":"10.1016/j.mrfmmm.2025.111912","url":null,"abstract":"<div><h3>Background</h3><div>Prostate cancer (PCa) incidence increases as age advances and seriously endangers men’s health worldwide. Arginase 2 (ARG2) has been identified as a potential diagnostic and prognostic marker for PCa. However, the molecular mechanisms underlying its function in PCa remain undefined.</div></div><div><h3>Methods</h3><div>ARG2 mRNA and protein expression were quantified in PCa tissues and cells using qRT-PCR and Western blot. Cellular proliferation, glucose consumption, lactate production, apoptosis, and ferroptosis were evaluated via EdU incorporation, colony formation assays, commercial kits, and flow cytometry. Subsequently, the xenograft model was established to assess ARG2’s role in tumor growth <em>in vivo</em>. Bioinformatics analysis and RNA immunoprecipitation (RIP) were employed to investigate the interaction between Wilms’ tumor 1-associating protein (WTAP), a key component of the N6-methyladenosine (m6A) methyltransferase complex, and ARG2 mRNA. Besides, mRNA stability was determined using actinomycin D chase assays.</div></div><div><h3>Results</h3><div>ARG2 exhibited low expression in PCa tissues and cells. Upregulation of ARG2 inhibited proliferation and glycolysis, and promoted apoptosis, oxidative stress and ferroptosis of PCa cells. However, silencing ARG2 had the opposite effects. I<em>n vivo</em>, ARG2 overexpression suppressed tumor growth. Mechanistically, WTAP bound directly to ARG2 mRNA, and their expression levels were inversely correlated. WTAP knockdown phenocopied ARG2 overexpression by repressing proliferation and glycolysis and enhancing apoptosis/ferroptosis, effects reversed by ARG2 silencing. ARG2 overexpression counteracted the oncogenic effects of WTAP overexpression.</div></div><div><h3>Conclusion</h3><div>WTAP bound to ARG2 and suppressed its expression, thereby promoting the malignant progression of PCa.</div></div>","PeriodicalId":49790,"journal":{"name":"Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis","volume":"831 ","pages":"Article 111912"},"PeriodicalIF":1.9,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144750383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

DGKβ accelerates the progression of cervical cancer through ANGPT4-mediated tumor angiogenesis DGKβ通过angpt4介导的肿瘤血管生成加速宫颈癌的进展

IF 1.9 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis

Pub Date : 2025-07-01 DOI: 10.1016/j.mrfmmm.2025.111913

Qing Li, Zhenyu Zhou, Xiaoying Li, Qiongyu Lan

Background

Cervical cancer (CC) is a major cause of morbidity and mortality in women, with complex etiology and progression. Diacylglycerol kinases (DGKs) are pivotal in lipid metabolism. Although diacylglycerol kinase beta (DGKβ) is well-studied in neurology, its role in cancer, especially CC, remains underexplored. This study aimed to explore DGKβ's role and mechanism in CC.

Methods

Bioinformatics analysis was employed to identify genes differentially expressed in CC, with western blot confirming DGKβ expression in CC cells. The role of DGKβ was examined through small interfering RNA-mediated gene silencing, proliferation tests, migration and invasion assays, and angiogenesis studies. In-depth bioinformatics explored DGKβ-regulated downstream targets and pathways. Pathological assessment elucidated the impact of DGKβ and angiopoietin 4 (ANGPT4) on CC samples.

Results

Our data identified DGKβ as a promising candidate gene in the context of CC. This conclusion stemmed from the notable observation that DGKβ exhibited a heightened expression in CC cell lines. Notably, the silencing of DGKβ resulted in the suppression of CC cell proliferation, invasion, migration, as well as the epithelial-mesenchymal transition processes. Additional bioinformatics analysis delving into DGKβ-associated genes revealed ANGPT4 as a downstream target gene of DGKβ, which is capable of modulating angiogenesis and possesses multiple cellular functions related to cell survival, proliferation, and migration. Most significantly, our findings also demonstrated that both DGKβ and ANGPT4 were overexpressed in clinical specimens of CC.

Conclusion

This study uncovered an oncogenic role for DGKβ in CC and identified a potential regulatory link between DGKβ and ANGPT4 in tumor angiogenesis. These findings provided promising directions for developing new diagnostic and therapeutic approaches for CC.

宫颈癌（CC）是女性发病和死亡的主要原因，其病因和进展复杂。二酰基甘油激酶（DGKs）在脂质代谢中起关键作用。尽管二酰基甘油激酶β (DGKβ)在神经学中得到了很好的研究，但其在癌症，特别是CC中的作用仍未得到充分研究。目的探讨DGKβ在CC中的作用及机制。方法采用生物信息学方法鉴定CC细胞中DGKβ的差异表达基因，western blot证实DGKβ在CC细胞中的表达。DGKβ的作用通过小干扰rna介导的基因沉默、增殖试验、迁移和侵袭试验以及血管生成研究来检测。深入的生物信息学探索dgk β调控的下游靶点和途径。病理评估阐明DGKβ和血管生成素4 （ANGPT4）对CC样品的影响。结果DGKβ在CC细胞系中表达增高，这一结论来源于DGKβ在CC细胞系中的显著表达。值得注意的是，DGKβ的沉默导致CC细胞增殖、侵袭、迁移以及上皮-间质转化过程的抑制。进一步深入DGKβ相关基因的生物信息学分析表明，ANGPT4是DGKβ的下游靶基因，能够调节血管生成，并具有与细胞存活、增殖和迁移相关的多种细胞功能。最重要的是，我们的研究结果还表明DGKβ和ANGPT4在CC的临床标本中都过表达。结论本研究揭示了DGKβ在CC中的致癌作用，并确定了DGKβ和ANGPT4在肿瘤血管生成中的潜在调节联系。这些发现为开发新的CC诊断和治疗方法提供了有希望的方向。

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引用次数: 0