KDM5B is a histone H3K4me2/3 demethylase. The PHD1 domain of KDM5B is critical for demethylation, but the mechanism underlying the action of this domain is unclear. In this paper, we observed that PHDIKDMSB interacts ...KDM5B is a histone H3K4me2/3 demethylase. The PHD1 domain of KDM5B is critical for demethylation, but the mechanism underlying the action of this domain is unclear. In this paper, we observed that PHDIKDMSB interacts with unmethylated H3K4me0. Our NMR structure of PHDIKDMSB in complex with H3K4me0 revealed that the binding mode is slightly different from that of other reported PHD fingers. The disruption of this interaction by double mutations on the residues in the interface （L325A/D328A） decreases the H3K4me2/3 demethylation activity of KDM5B in cells by approximately 50% and increases the transcriptional repression of tumor suppressor genes by approximately twofold. These findings imply that PHDIKDMSB may help maintain KDM5B at target genes to mediate the demethylation activities of KDM5B.展开更多
BACKGROUND Colorectal cancer(CRC)is one of the most common causes of cancer mortality worldwide.The transcription factor Myc-associated zinc finger protein(MAZ)has been implicated in cancer progression.However,its pre...BACKGROUND Colorectal cancer(CRC)is one of the most common causes of cancer mortality worldwide.The transcription factor Myc-associated zinc finger protein(MAZ)has been implicated in cancer progression.However,its precise function and mecha-nisms in CRC remain unclear.AIM To investigate the role and mechanism of the MAZ/ubiquitin-like with PHD and RING finger domains 1(UHRF1)/esophageal cancer-related gene 4(ECRG4)axis in CRC metastasis.METHODS Western blot,quantitative reverse transcription polymerase chain reaction(PCR)and transwell were performed to evaluate the impact of MAZ knockdown on CRC cell migration and invasion.A xenograft tumor metastasis model was es-tablished by injecting MAZ-deficient CRC cells into nude mice to assess in vivo metastatic potential.Dual-luciferase reporter assay was performed to determine the role of MAZ and its downstream target,UHRF1.Chromatin immunoprecip-itation-quantitative PCR and methylation-specific PCR were used to analyze whether UHRF1 regulated ECRG4 through DNA methylation.RESULTS MAZ was highly upregulated in CRC cells and promoted CRC migration,inva-sion,epithelial-mesenchymal transition(EMT)and metastasis.Mechanistically,MAZ transcriptionally activated UHRF1,which in turn led to DNA methylation of ECRG4.Knockdown of MAZ suppressed CRC migration and invasion was reversed by overexpression of UHRF1.Loss of UHRF1 upregulated ECRG4,inhibited EMT,and reduced cell migration and invasion.However,simultaneous knockdown of ECRG4 partially reversed these effects.CONCLUSION MAZ promotes CRC cell migration,invasion,and EMT by transcriptionally activating UHRF1,which downreg-ulates ECRG4 through DNA methylation.展开更多
利用生物信息学数据库分析泛素样含PHD和环指结构域蛋白1(UHRF1)在恶性胸膜间皮瘤(MPM)中的表达水平及临床意义。基于TCGA数据库和GTEx数据库差异表达分析UHRF1 m RNA在MPM组织和正常肺组织中的表达水平;使用R软件分析UHRF1 mRNA表达量...利用生物信息学数据库分析泛素样含PHD和环指结构域蛋白1(UHRF1)在恶性胸膜间皮瘤(MPM)中的表达水平及临床意义。基于TCGA数据库和GTEx数据库差异表达分析UHRF1 m RNA在MPM组织和正常肺组织中的表达水平;使用R软件分析UHRF1 mRNA表达量与临床病理参数的相关性;构建Kaplan-Meier模型和单因素多因素COX回归模型分析UHRF1基因在MPM中的预后;利用TIMER2.0数据库分析UHRF1基因与免疫细胞浸润的关系;GSEA分析UHRF1基因发挥功能的主要富集通路。选取8例MPM组织及4例非MPM胸膜组织,通过RT-q PCR的方法验证UHRF1在MPM与非MPM胸膜组织的表达情况。数据库分析结果表明,与正常肺组织相比,UHRF1 m RNA在MPM组织中高表达;UHRF1高表达患者提示MPM患者预后不良;UHRF1基因表达量与CD4^(+)辅助型T细胞2、CD4^(+)效应记忆性T细胞、巨噬细胞等多种免疫细胞浸润水平具有显著的相关性(P<0.01),且显著影响MPM患者的预后。功能富集分析显示,UHRF1主要在DNA复制、蛋白酶体、同源重组等通路中起作用。在收集到的病例样本中,与非MPM胸膜组织相比,UHRF1 mRNA在MPM组织中的表达显著增高(P<0.001)。UHRF1在MPM组织中高表达,可能通过调节DNA甲基化和免疫细胞浸润来影响MPM患者预后,有望成为MPM治疗和预后评估的潜在靶点。展开更多
泛素样含PHD和环指域1(ubiquitin-like with PHD and ring finger domains 1,UHRF1)是新发现的一种与细胞生长有关的核蛋白基因,参与了细胞增殖、周期调控、凋亡及放射敏感性等重要的生物学过程。并且在多种肿瘤细胞中异常高表达,与肿...泛素样含PHD和环指域1(ubiquitin-like with PHD and ring finger domains 1,UHRF1)是新发现的一种与细胞生长有关的核蛋白基因,参与了细胞增殖、周期调控、凋亡及放射敏感性等重要的生物学过程。并且在多种肿瘤细胞中异常高表达,与肿瘤的发生、发展、侵袭转移及放化疗抵抗关系密切,可能成为肿瘤基因治疗的一个新靶点。本文就UHRF1与肿瘤的关系作一简要综述。展开更多
目的探讨下调泛素样含PHD和环指域1(ubiquitin-like protein containing PHD and RING finger domains 1,UHRF1)基因在喉癌Hep-2细胞中的表达后对其增殖,凋亡,侵袭迁移能力的影响。方法实验分为空白组,阴性对照组和干扰组。利用UHRF1-si...目的探讨下调泛素样含PHD和环指域1(ubiquitin-like protein containing PHD and RING finger domains 1,UHRF1)基因在喉癌Hep-2细胞中的表达后对其增殖,凋亡,侵袭迁移能力的影响。方法实验分为空白组,阴性对照组和干扰组。利用UHRF1-si RNA干扰UHRF1在喉癌Hep-2细胞中的表达,采用q RT-PCR、Western blot检测UHRF1的表达变化。采用MTT法、流式细胞术、Transwell实验检测下调UHRF1表达后Hep-2细胞的增殖、凋亡、侵袭、迁移能力的变化。Western blot检测下调UHRF1表达后Hep-2细胞内Bax、Bcl-2、MMP-2、MMP-9蛋白表达的变化情况。结果与空白组和阴性对照组相比干扰组下调UHRF1的表达后可明显抑制喉癌Hep-2细胞的增殖能力(P<0.001)。空白组,阴性对照组,干扰组凋亡率分别为(5.47±2.77)%,(3.95±0.66)%,(18.95±1.10)%,干扰组凋亡率明显提高(P<0.001)。空白组,阴性对照,干扰组每视野侵袭细胞数分别为(213.7±13.1),(221.3±10.3),(97.7±7.5),干扰组侵袭细胞数明显降低(P<0.001)。空白组,阴性对照,干扰组每视野迁移细胞数分别为(230.7±5.5),(222.7±11.2),(111.7±7.6),干扰组迁移细胞数明显降低(P<0.001)。干扰组细胞Bax蛋白表达上调(P<0.001),Bcl-2蛋白表达下调(P<0.001),MMP-2蛋白、MMP-9蛋白表达下调(P<0.001)。结论在喉癌Hep-2细胞中下调UHRF1的表达后可明显抑制细胞的增殖,促进细胞凋亡,抑制细胞的侵袭迁移能力。展开更多
文摘KDM5B is a histone H3K4me2/3 demethylase. The PHD1 domain of KDM5B is critical for demethylation, but the mechanism underlying the action of this domain is unclear. In this paper, we observed that PHDIKDMSB interacts with unmethylated H3K4me0. Our NMR structure of PHDIKDMSB in complex with H3K4me0 revealed that the binding mode is slightly different from that of other reported PHD fingers. The disruption of this interaction by double mutations on the residues in the interface （L325A/D328A） decreases the H3K4me2/3 demethylation activity of KDM5B in cells by approximately 50% and increases the transcriptional repression of tumor suppressor genes by approximately twofold. These findings imply that PHDIKDMSB may help maintain KDM5B at target genes to mediate the demethylation activities of KDM5B.
基金Supported by Hangzhou Medical and Health Science and Technology Plan,No.B20210014.
文摘BACKGROUND Colorectal cancer(CRC)is one of the most common causes of cancer mortality worldwide.The transcription factor Myc-associated zinc finger protein(MAZ)has been implicated in cancer progression.However,its precise function and mecha-nisms in CRC remain unclear.AIM To investigate the role and mechanism of the MAZ/ubiquitin-like with PHD and RING finger domains 1(UHRF1)/esophageal cancer-related gene 4(ECRG4)axis in CRC metastasis.METHODS Western blot,quantitative reverse transcription polymerase chain reaction(PCR)and transwell were performed to evaluate the impact of MAZ knockdown on CRC cell migration and invasion.A xenograft tumor metastasis model was es-tablished by injecting MAZ-deficient CRC cells into nude mice to assess in vivo metastatic potential.Dual-luciferase reporter assay was performed to determine the role of MAZ and its downstream target,UHRF1.Chromatin immunoprecip-itation-quantitative PCR and methylation-specific PCR were used to analyze whether UHRF1 regulated ECRG4 through DNA methylation.RESULTS MAZ was highly upregulated in CRC cells and promoted CRC migration,inva-sion,epithelial-mesenchymal transition(EMT)and metastasis.Mechanistically,MAZ transcriptionally activated UHRF1,which in turn led to DNA methylation of ECRG4.Knockdown of MAZ suppressed CRC migration and invasion was reversed by overexpression of UHRF1.Loss of UHRF1 upregulated ECRG4,inhibited EMT,and reduced cell migration and invasion.However,simultaneous knockdown of ECRG4 partially reversed these effects.CONCLUSION MAZ promotes CRC cell migration,invasion,and EMT by transcriptionally activating UHRF1,which downreg-ulates ECRG4 through DNA methylation.
文摘利用生物信息学数据库分析泛素样含PHD和环指结构域蛋白1(UHRF1)在恶性胸膜间皮瘤(MPM)中的表达水平及临床意义。基于TCGA数据库和GTEx数据库差异表达分析UHRF1 m RNA在MPM组织和正常肺组织中的表达水平;使用R软件分析UHRF1 mRNA表达量与临床病理参数的相关性;构建Kaplan-Meier模型和单因素多因素COX回归模型分析UHRF1基因在MPM中的预后;利用TIMER2.0数据库分析UHRF1基因与免疫细胞浸润的关系;GSEA分析UHRF1基因发挥功能的主要富集通路。选取8例MPM组织及4例非MPM胸膜组织,通过RT-q PCR的方法验证UHRF1在MPM与非MPM胸膜组织的表达情况。数据库分析结果表明,与正常肺组织相比,UHRF1 m RNA在MPM组织中高表达;UHRF1高表达患者提示MPM患者预后不良;UHRF1基因表达量与CD4^(+)辅助型T细胞2、CD4^(+)效应记忆性T细胞、巨噬细胞等多种免疫细胞浸润水平具有显著的相关性(P<0.01),且显著影响MPM患者的预后。功能富集分析显示,UHRF1主要在DNA复制、蛋白酶体、同源重组等通路中起作用。在收集到的病例样本中,与非MPM胸膜组织相比,UHRF1 mRNA在MPM组织中的表达显著增高(P<0.001)。UHRF1在MPM组织中高表达,可能通过调节DNA甲基化和免疫细胞浸润来影响MPM患者预后,有望成为MPM治疗和预后评估的潜在靶点。
文摘泛素样含PHD和环指域1(ubiquitin-like with PHD and ring finger domains 1,UHRF1)是新发现的一种与细胞生长有关的核蛋白基因,参与了细胞增殖、周期调控、凋亡及放射敏感性等重要的生物学过程。并且在多种肿瘤细胞中异常高表达,与肿瘤的发生、发展、侵袭转移及放化疗抵抗关系密切,可能成为肿瘤基因治疗的一个新靶点。本文就UHRF1与肿瘤的关系作一简要综述。
文摘目的探讨下调泛素样含PHD和环指域1(ubiquitin-like protein containing PHD and RING finger domains 1,UHRF1)基因在喉癌Hep-2细胞中的表达后对其增殖,凋亡,侵袭迁移能力的影响。方法实验分为空白组,阴性对照组和干扰组。利用UHRF1-si RNA干扰UHRF1在喉癌Hep-2细胞中的表达,采用q RT-PCR、Western blot检测UHRF1的表达变化。采用MTT法、流式细胞术、Transwell实验检测下调UHRF1表达后Hep-2细胞的增殖、凋亡、侵袭、迁移能力的变化。Western blot检测下调UHRF1表达后Hep-2细胞内Bax、Bcl-2、MMP-2、MMP-9蛋白表达的变化情况。结果与空白组和阴性对照组相比干扰组下调UHRF1的表达后可明显抑制喉癌Hep-2细胞的增殖能力(P<0.001)。空白组,阴性对照组,干扰组凋亡率分别为(5.47±2.77)%,(3.95±0.66)%,(18.95±1.10)%,干扰组凋亡率明显提高(P<0.001)。空白组,阴性对照,干扰组每视野侵袭细胞数分别为(213.7±13.1),(221.3±10.3),(97.7±7.5),干扰组侵袭细胞数明显降低(P<0.001)。空白组,阴性对照,干扰组每视野迁移细胞数分别为(230.7±5.5),(222.7±11.2),(111.7±7.6),干扰组迁移细胞数明显降低(P<0.001)。干扰组细胞Bax蛋白表达上调(P<0.001),Bcl-2蛋白表达下调(P<0.001),MMP-2蛋白、MMP-9蛋白表达下调(P<0.001)。结论在喉癌Hep-2细胞中下调UHRF1的表达后可明显抑制细胞的增殖,促进细胞凋亡,抑制细胞的侵袭迁移能力。
