目的探讨环指蛋白2(ring finger protein 2,RNF2)在胃腺癌组织中的表达及临床意义。方法采用免疫组化EnVision法检测80例胃腺癌及癌旁组织中RNF2蛋白表达水平,分析RNF2蛋白表达与胃腺癌临床病理特征及预后的关系。RT-qPCR法分析RNF2 mRN...目的探讨环指蛋白2(ring finger protein 2,RNF2)在胃腺癌组织中的表达及临床意义。方法采用免疫组化EnVision法检测80例胃腺癌及癌旁组织中RNF2蛋白表达水平,分析RNF2蛋白表达与胃腺癌临床病理特征及预后的关系。RT-qPCR法分析RNF2 mRNA在11例胃腺癌和癌旁组织中的表达水平。采用Spearman相关性分析RNF2与E-Cadherin蛋白表达的相关性。结果RNF2蛋白在胃腺癌组织(76.3%)中的表达明显高于癌旁组织(25.0%),差异有统计学意义(P﹤0.05)。RNF2表达与胃腺癌肿瘤大小、浸润深度、淋巴结转移、TNM分期及预后相关(P均﹤0.05),与患者性别、年龄、肿瘤分化程度、Lauren分型及神经侵犯无关(P﹥0.05)。RT-qPCR检测结果显示,RNF2 mRNA在胃腺癌组织(11.92±3.48)中的水平显著高于癌旁组织(11.11±2.38)(P﹤0.05)。相关性分析显示,RNF2与E-Cadherin在胃腺癌组织中的表达呈负相关(r_(s)=-0.278,P﹤0.05)。结论RNF2在胃腺癌的发生、发展中具有促进作用,表明RNF2可能成为胃腺癌治疗的潜在靶点。展开更多
为分析E3泛素连接酶环指蛋白125(ring finger protein 125,RNF125)在肝癌中的表达及临床预后意义,并探究RNF125对肝癌侵袭和转移的影响及其作用机制,通过ENCORI数据库和临床组织样本分析RNF125在肝癌和癌旁组织中的表达量,使用ENCORI数...为分析E3泛素连接酶环指蛋白125(ring finger protein 125,RNF125)在肝癌中的表达及临床预后意义,并探究RNF125对肝癌侵袭和转移的影响及其作用机制,通过ENCORI数据库和临床组织样本分析RNF125在肝癌和癌旁组织中的表达量,使用ENCORI数据库分析RNF125与肝癌预后的关系。构建敲低和过表达RNF125的肝癌细胞系,通过Transwell试验和细胞划痕试验检验RNF125表达水平对肝癌细胞迁移、侵袭能力的影响。通过动物试验验证调控RNF125对体内肿瘤生长情况及对肿瘤免疫微环境的影响,质谱分析寻找RNF125的靶蛋白,qPCR、蛋白质免疫印迹技术、泛素化试验验证RNF125与MYH9之间的关系,通过蛋白质免疫印迹和细胞免疫荧光技术确定调控RNF125与MYH9对EMT标志蛋白的影响。结果表明:RNF125在肝癌组织中表达较低,RNF125的高表达提示良好的预后。RNF125的表达水平与肝癌细胞迁移和侵袭能力呈负相关。过表达RNF125使肿瘤生长延迟并增加免疫细胞的浸润水平。RNF125特异性结合MYH9并促进其泛素化降解。RNF125与上皮细胞标志物的表达水平呈正相关,与间充质细胞标志物呈负相关。过表达MYH9可逆转RNF125对EMT标志蛋白的影响。综上,RNF125在肝癌中表达下调,RNF125的高表达与肝癌患者的良好预后相关,RNF125通过泛素化MYH9来抑制肝癌的EMT过程,从而抑制肝癌的迁移和侵袭。展开更多
口蹄疫病毒(FMDV)编码四种结构蛋白,其中VP1蛋白是病毒表面的主要衣壳蛋白,在病毒入侵和抗原性方面发挥关键作用,决定病毒的致病力和组织嗜性。2025年1月17日,中国农业科学院兰州兽医研究所郑海学研究员带领的科研团队在PLoS Pathogens...口蹄疫病毒(FMDV)编码四种结构蛋白,其中VP1蛋白是病毒表面的主要衣壳蛋白,在病毒入侵和抗原性方面发挥关键作用,决定病毒的致病力和组织嗜性。2025年1月17日,中国农业科学院兰州兽医研究所郑海学研究员带领的科研团队在PLoS Pathogens期刊上发表了题为“RING finger protein 5 is a key anti-FMDV host factor through inhibition of virion assembly”的研究论文,该研究揭示了环指蛋白5(RNF5)依赖其E3泛素酶活性介导FMDV VP1蛋白降解并限制病毒组装的功能机制,为广谱抗小RNA病毒药物的研发提供了理论依据,为抗病育种提供新思路。展开更多
Ring finger protein 122(RNF122),an E3 ubiquitin ligase,orchestrates antiviral immune responses in mammals by targeting retinoic acid-inducible gene 1 and melanoma differentiation-associated gene 5 for ubiquitination.H...Ring finger protein 122(RNF122),an E3 ubiquitin ligase,orchestrates antiviral immune responses in mammals by targeting retinoic acid-inducible gene 1 and melanoma differentiation-associated gene 5 for ubiquitination.However,its functional relevance in teleosts has yet to be clearly defined,particularly regarding the identification of substrate-specific regulatory sites.This study characterized RNF122 from mandarin fish(Siniperca chuatsi),termed scRNF122,and investigated its regulatory impact on stimulator of interferon genes(STING)-mediated antiviral signaling.Results showed that scRNF122 expression was up-regulated in response to mandarin fish ranavirus(MRV)infection,and its overexpression suppressed scSTING-mediated interferon(IFN)production and enhanced MRV replication.Co-immunoprecipitation confirmed a direct interaction between scRNF122 and scSTING.Functional assays demonstrated that scRNF122 facilitated scSTING degradation through the ubiquitin-proteasome pathway,a process impeded by MG132 treatment.Ubiquitination analyses of various scSTING mutants revealed that scRNF122 catalyzed scSTING ubiquitination at K95,K117,and K155 residues.Moreover,scRNF122 significantly impaired scSTING-dependent antiviral responses by engaging negative regulatory elements within the signaling cascade.Overall,scRNF122 was identified as a negative modulator of STING-mediated IFN signaling in mandarin fish,diminishing STING-dependent antiviral activity and promoting its degradation via the ubiquitin-proteasome pathway at lysine residues K95,K117,and K155.These findings provide mechanistic insight into the post-translational control of STING in teleosts and establish a foundation for future investigations into antiviral immune regulation.展开更多
目的利用慢病毒干涉下调内源性RNF31表达,研究NF-κB通路的活化及对细胞凋亡的影响。方法将人RNF31的shRNA片段克隆到慢病毒表达载体p Green Puro中,瞬时转染HEK293T细胞,筛选出有效的干涉片段。将重组表达质粒与包装质粒PMD、SPA共转染...目的利用慢病毒干涉下调内源性RNF31表达,研究NF-κB通路的活化及对细胞凋亡的影响。方法将人RNF31的shRNA片段克隆到慢病毒表达载体p Green Puro中,瞬时转染HEK293T细胞,筛选出有效的干涉片段。将重组表达质粒与包装质粒PMD、SPA共转染293T细胞,在24 h、48 h分2次收集慢病毒上清,用流式细胞术检测病毒滴度。将获得的病毒感染HEK293细胞,提取细胞蛋白,Real-time PCR以及Western Blot检测RNF31干涉效果;报告基因实验检测敲低RNF31对NF-κB转录活性的影响;Real-time PCR检测干涉RNF31对TNF-α诱导的NF-κB下游靶基因的影响;Western Blot检测下调RNF31对IκBα活化的影响;Hochest染色检测下调RNF31对细胞凋亡的影响。结果成功构建RNF31干涉慢病毒p Green Puro-RNF31载体并获得慢病毒颗粒,病毒滴度可达3×107pfu/ml。在HEK293细胞中下调RNF31,抑制TNF-α刺激的NF-κB的转录活性,并抑制NF-κB下游靶基因的表达;下调RNF31抑制TNF-α刺激的IκBα的活化;此外,在TNF-α刺激细胞24 h时,RNF31表达下调使细胞凋亡增多。结论 RNF31表达下调抑制TNF-α刺激的NF-κB通路的激活。展开更多
文摘为分析E3泛素连接酶环指蛋白125(ring finger protein 125,RNF125)在肝癌中的表达及临床预后意义,并探究RNF125对肝癌侵袭和转移的影响及其作用机制,通过ENCORI数据库和临床组织样本分析RNF125在肝癌和癌旁组织中的表达量,使用ENCORI数据库分析RNF125与肝癌预后的关系。构建敲低和过表达RNF125的肝癌细胞系,通过Transwell试验和细胞划痕试验检验RNF125表达水平对肝癌细胞迁移、侵袭能力的影响。通过动物试验验证调控RNF125对体内肿瘤生长情况及对肿瘤免疫微环境的影响,质谱分析寻找RNF125的靶蛋白,qPCR、蛋白质免疫印迹技术、泛素化试验验证RNF125与MYH9之间的关系,通过蛋白质免疫印迹和细胞免疫荧光技术确定调控RNF125与MYH9对EMT标志蛋白的影响。结果表明:RNF125在肝癌组织中表达较低,RNF125的高表达提示良好的预后。RNF125的表达水平与肝癌细胞迁移和侵袭能力呈负相关。过表达RNF125使肿瘤生长延迟并增加免疫细胞的浸润水平。RNF125特异性结合MYH9并促进其泛素化降解。RNF125与上皮细胞标志物的表达水平呈正相关,与间充质细胞标志物呈负相关。过表达MYH9可逆转RNF125对EMT标志蛋白的影响。综上,RNF125在肝癌中表达下调,RNF125的高表达与肝癌患者的良好预后相关,RNF125通过泛素化MYH9来抑制肝癌的EMT过程,从而抑制肝癌的迁移和侵袭。
文摘口蹄疫病毒(FMDV)编码四种结构蛋白,其中VP1蛋白是病毒表面的主要衣壳蛋白,在病毒入侵和抗原性方面发挥关键作用,决定病毒的致病力和组织嗜性。2025年1月17日,中国农业科学院兰州兽医研究所郑海学研究员带领的科研团队在PLoS Pathogens期刊上发表了题为“RING finger protein 5 is a key anti-FMDV host factor through inhibition of virion assembly”的研究论文,该研究揭示了环指蛋白5(RNF5)依赖其E3泛素酶活性介导FMDV VP1蛋白降解并限制病毒组装的功能机制,为广谱抗小RNA病毒药物的研发提供了理论依据,为抗病育种提供新思路。
基金supported by the National Key Research and Development Program of China(2022YFE0203900,2024YFD2401101)China Agriculture Research System(CARS-46)+1 种基金National Natural Science Foundation of China(32473201)Guangdong S&T Program(2022B1111030001,2024B1212040007)。
文摘Ring finger protein 122(RNF122),an E3 ubiquitin ligase,orchestrates antiviral immune responses in mammals by targeting retinoic acid-inducible gene 1 and melanoma differentiation-associated gene 5 for ubiquitination.However,its functional relevance in teleosts has yet to be clearly defined,particularly regarding the identification of substrate-specific regulatory sites.This study characterized RNF122 from mandarin fish(Siniperca chuatsi),termed scRNF122,and investigated its regulatory impact on stimulator of interferon genes(STING)-mediated antiviral signaling.Results showed that scRNF122 expression was up-regulated in response to mandarin fish ranavirus(MRV)infection,and its overexpression suppressed scSTING-mediated interferon(IFN)production and enhanced MRV replication.Co-immunoprecipitation confirmed a direct interaction between scRNF122 and scSTING.Functional assays demonstrated that scRNF122 facilitated scSTING degradation through the ubiquitin-proteasome pathway,a process impeded by MG132 treatment.Ubiquitination analyses of various scSTING mutants revealed that scRNF122 catalyzed scSTING ubiquitination at K95,K117,and K155 residues.Moreover,scRNF122 significantly impaired scSTING-dependent antiviral responses by engaging negative regulatory elements within the signaling cascade.Overall,scRNF122 was identified as a negative modulator of STING-mediated IFN signaling in mandarin fish,diminishing STING-dependent antiviral activity and promoting its degradation via the ubiquitin-proteasome pathway at lysine residues K95,K117,and K155.These findings provide mechanistic insight into the post-translational control of STING in teleosts and establish a foundation for future investigations into antiviral immune regulation.
