目的:探讨乳酸对三阴性乳腺癌(triple-negative breast cancer,TNBC)细胞恶性行为的影响,并揭示胰岛素样生长因子2 mRNA结合蛋白3(insulin-like growth factor 2 m RNA binding protein 3,IGF2BP3)通过m6A依赖性方式调控EP300,进而介导...目的:探讨乳酸对三阴性乳腺癌(triple-negative breast cancer,TNBC)细胞恶性行为的影响,并揭示胰岛素样生长因子2 mRNA结合蛋白3(insulin-like growth factor 2 m RNA binding protein 3,IGF2BP3)通过m6A依赖性方式调控EP300,进而介导代谢重编程的表观遗传学机制。方法:通过外源性添加乳酸钠处理TNBC细胞,采用CCK-8、集落形成、划痕愈合和Transwell实验检测细胞增殖与迁移能力;利用shRNA慢病毒载体构建IGF2BP3稳定敲低细胞模型,结合Western blot分析组蛋白乳酸化修饰水平;进一步通过RIP-seq、MeRIP-seq筛选IGF2BP3的m6A靶基因,并利用qRT-PCR和Western blot验证关键分子表达。结果:乳酸钠处理显著增强了TNBC细胞的增殖和迁移能力,且泛乳酸化水平随乳酸浓度梯度升高而增加。IGF2BP3敲低可降低组蛋白乳酸化修饰水平。通过整合RIP-seq和MeRIP-seq数据,鉴定出699个其转录本同时被IGF2BP3特异性结合且携带m6A修饰的候选基因,这些基因显著富集于表观遗传调控和代谢相关通路。EP300转录本上IGF2BP3的结合区域与m6A修饰位点高度重合,且EP300的表达水平在IGF2BP3敲低后明显下调。结论:外源性乳酸通过泛乳酸化修饰促进TNBC的恶性进展。IGF2BP3通过m6A依赖性机制调控EP300的表达,进而介导组蛋白乳酸化修饰与代谢重编程的协同作用,最终驱动TNBC的肿瘤进展。上述发现为靶向代谢-表观遗传交互调控的新型治疗策略提供了理论依据。展开更多
Numerous studies have characterized the critical role of circular RNAs(circRNAs)as regulatory factors in the progression of multiple cancers.However,the biological functions of circRNAs and their underlying molecular ...Numerous studies have characterized the critical role of circular RNAs(circRNAs)as regulatory factors in the progression of multiple cancers.However,the biological functions of circRNAs and their underlying molecular mechanisms in the progression of uveal melanoma(UM)remain enigmatic.In this study,we identified a novel circRNA,circ_0053943,through re-analysis of UM microarray data and quantitative RT-PCR.Circ_0053943 was found to be upregulated in UM and to promote the proliferation and metastatic ability of UM cells in both in vitro and in vivo settings.Mechanistically,circ_0053943 was observed to bind to the KH1 and KH2 domains of insulin-like growth factor 2 mRNA-binding protein 3(IGF2BP3),thereby enhancing the function of IGF2BP3 by stabilizing its target mRNA.RNA sequencing assays identified epidermal growth factor receptor(EGFR)as a target gene of circ_0053943 and IGF2BP3 at the transcriptional level.Rescue assays demonstrated that circ_0053943 exerts its biological function by stabilizing EGFR mRNA and regulating the downstream mitogen-activated protein kinase/extracellular signal-regulated kinase(MAPK/ERK)signaling pathway.Collectively,circ_0053943 may promote UM progression by stabilizing EGFR mRNA and activating the MAPK/ERK signaling pathway through the formation of a circ_0053943/IGF2BP3/EGFR RNA-protein ternary complex,thus providing a potential biomarker and therapeutic target for UM.展开更多
Objective Gastric cancer(GC)is one of the most prevalent cancers worldwide and is associated with high morbidity and mortality rates.The IGF2 mRNA-binding protein(IGF2BP)participates in a variety of cancers.The aim of...Objective Gastric cancer(GC)is one of the most prevalent cancers worldwide and is associated with high morbidity and mortality rates.The IGF2 mRNA-binding protein(IGF2BP)participates in a variety of cancers.The aim of this study was to analyze the expression of IGF2BP3 and explore the genes related to IGF2BP3 in GC.Methods Bioinformatics software was used to analyze the expression of IGF2BP1,IGF2BP2,and IGF2BP3 in tumors,and the expression of IGF2BPs in the GSE118897 dataset.Immunohistochemistry was performed to detect the protein level of IGF2BP3 in GC samples.cBioPortal was used to query gene alteration of IGF2BP3.LinkedOmics was used to identify genes related to IGF2BP3.Results Sangerbox analysis showed that the expression of all IGF2BP family members was higher in GC.cBioporta analysis showed that gene alteration of IGF2BP3 in stomach adenocarcinoma included mutation and amplificatio.LinkedOmics analysis showed that many genes were correlated with IGF2BP3,such as PLAGL2,GET4,IGF2BP1,HMGA2,CLDN6,HOXC13,SMARCA2,TMEM66,CIRBP,NFIX,SLC25A12,and CYB5D2.Conclusion In this study,we founded that IGF2BP3 was overexpressed in GC.Furthermore,this study identified potential genes related to IGF2BP3 in GC,which should be studied further.展开更多
文摘目的:探讨乳酸对三阴性乳腺癌(triple-negative breast cancer,TNBC)细胞恶性行为的影响,并揭示胰岛素样生长因子2 mRNA结合蛋白3(insulin-like growth factor 2 m RNA binding protein 3,IGF2BP3)通过m6A依赖性方式调控EP300,进而介导代谢重编程的表观遗传学机制。方法:通过外源性添加乳酸钠处理TNBC细胞,采用CCK-8、集落形成、划痕愈合和Transwell实验检测细胞增殖与迁移能力;利用shRNA慢病毒载体构建IGF2BP3稳定敲低细胞模型,结合Western blot分析组蛋白乳酸化修饰水平;进一步通过RIP-seq、MeRIP-seq筛选IGF2BP3的m6A靶基因,并利用qRT-PCR和Western blot验证关键分子表达。结果:乳酸钠处理显著增强了TNBC细胞的增殖和迁移能力,且泛乳酸化水平随乳酸浓度梯度升高而增加。IGF2BP3敲低可降低组蛋白乳酸化修饰水平。通过整合RIP-seq和MeRIP-seq数据,鉴定出699个其转录本同时被IGF2BP3特异性结合且携带m6A修饰的候选基因,这些基因显著富集于表观遗传调控和代谢相关通路。EP300转录本上IGF2BP3的结合区域与m6A修饰位点高度重合,且EP300的表达水平在IGF2BP3敲低后明显下调。结论:外源性乳酸通过泛乳酸化修饰促进TNBC的恶性进展。IGF2BP3通过m6A依赖性机制调控EP300的表达,进而介导组蛋白乳酸化修饰与代谢重编程的协同作用,最终驱动TNBC的肿瘤进展。上述发现为靶向代谢-表观遗传交互调控的新型治疗策略提供了理论依据。
基金国家自然科学基金(82270162,82270224,82070178)北京市自然科学基金(7222175)+2 种基金军队卫勤保障能力创新与生成专项(21WQ034)保健专项科研课题重点项目(21BJZ30)国家重点研发计划(2021Y FA 1100904)。
文摘目的:探寻IGF2BP3基因表达水平与急性髓系白血病(AML)患者预后的关系。方法:通过对本中心27例AML患者骨髓原代白血病细胞进行转录组高通量测序,分析IGF2BP3基因表达水平与患者临床特征之间的关系,并在初治AML患者及难治AML(Refractory AML)患者样本中验证。分析20例健康对照者和26例AML患者中IGF2BP3基因表达水平的差异。采用RT-qPCR、Western blot检测两种蒽环类耐药细胞系(HL60/ADR、K562/ADR)中IGF2BP3表达水平,比较其与敏感细胞(HL60、K562)的表达差异。通过3个数据集,分析IGF2BP3在AML患者中的表达水平及与预后的关系,进一步使用Cox生存分析IGF2BP3在AML中的预后价值。结果:在本中心27例AML患者骨髓原代白血病细胞中,难治性AML患者的IGF2BP3表达量明显高于化疗敏感的患者(P=0.0343),白血病细胞髓外浸润(extramedullary infiltration,EMI)患者的IGF2BP3表达量明显高于无髓外浸润的AML患者(P=0.0049)。与健康人比较,IGF2BP3在AML患者中表达增加(P=0.0009)。蒽环类耐药细胞系(HL60/ADR、K562/ADR)中IGF2BP3 mRNA的表达显著高于敏感细胞系(K562/ADR vs K562,P=0.0430;HL60/ADR vs HL60,P=0.7369)。Western blot结果显示,耐药细胞中IGF2BP3蛋白表达显著高于敏感细胞(P<0.001)。qPCR结果显示,难治AML患者中IGF2BP3的mRNA表达水平明显高于化疗敏感患者(P=0.002)。在3个大样本AML患者队列中IGF2BP3高表达预示AML预后不良(P<0.05)。单因素和多因素预后分析证实IGF2BP3高表达与患者较短的无事件生存(HR=1.887,P=0.024)和总体生存(HR=1.619,P=0.016)显著相关。结论:IGF2BP3基因高表达可能是AML预后不良的重要因素,提示IGF2BP3基因有望成为AML的临床预后评估和提供治疗策略的新的分子标志物。
基金supported by the National Natural Science Foundation of China(Nos.82273159 and 82171838)the Jiangsu Province’s Science and Technology Project(No.BE2020722).
文摘Numerous studies have characterized the critical role of circular RNAs(circRNAs)as regulatory factors in the progression of multiple cancers.However,the biological functions of circRNAs and their underlying molecular mechanisms in the progression of uveal melanoma(UM)remain enigmatic.In this study,we identified a novel circRNA,circ_0053943,through re-analysis of UM microarray data and quantitative RT-PCR.Circ_0053943 was found to be upregulated in UM and to promote the proliferation and metastatic ability of UM cells in both in vitro and in vivo settings.Mechanistically,circ_0053943 was observed to bind to the KH1 and KH2 domains of insulin-like growth factor 2 mRNA-binding protein 3(IGF2BP3),thereby enhancing the function of IGF2BP3 by stabilizing its target mRNA.RNA sequencing assays identified epidermal growth factor receptor(EGFR)as a target gene of circ_0053943 and IGF2BP3 at the transcriptional level.Rescue assays demonstrated that circ_0053943 exerts its biological function by stabilizing EGFR mRNA and regulating the downstream mitogen-activated protein kinase/extracellular signal-regulated kinase(MAPK/ERK)signaling pathway.Collectively,circ_0053943 may promote UM progression by stabilizing EGFR mRNA and activating the MAPK/ERK signaling pathway through the formation of a circ_0053943/IGF2BP3/EGFR RNA-protein ternary complex,thus providing a potential biomarker and therapeutic target for UM.
基金Supported by a grant from the National Natural Science Foundation of China(No.81802788)。
文摘Objective Gastric cancer(GC)is one of the most prevalent cancers worldwide and is associated with high morbidity and mortality rates.The IGF2 mRNA-binding protein(IGF2BP)participates in a variety of cancers.The aim of this study was to analyze the expression of IGF2BP3 and explore the genes related to IGF2BP3 in GC.Methods Bioinformatics software was used to analyze the expression of IGF2BP1,IGF2BP2,and IGF2BP3 in tumors,and the expression of IGF2BPs in the GSE118897 dataset.Immunohistochemistry was performed to detect the protein level of IGF2BP3 in GC samples.cBioPortal was used to query gene alteration of IGF2BP3.LinkedOmics was used to identify genes related to IGF2BP3.Results Sangerbox analysis showed that the expression of all IGF2BP family members was higher in GC.cBioporta analysis showed that gene alteration of IGF2BP3 in stomach adenocarcinoma included mutation and amplificatio.LinkedOmics analysis showed that many genes were correlated with IGF2BP3,such as PLAGL2,GET4,IGF2BP1,HMGA2,CLDN6,HOXC13,SMARCA2,TMEM66,CIRBP,NFIX,SLC25A12,and CYB5D2.Conclusion In this study,we founded that IGF2BP3 was overexpressed in GC.Furthermore,this study identified potential genes related to IGF2BP3 in GC,which should be studied further.
