目的通过生物信息学分析核受体亚家族6 A组成员1(nuclear receptor subfamily 6 group A member 1,NR6A1)在胃癌中的作用,旨在阐明NR6A1在胃癌中诊断和预后价值,也包括其与免疫细胞浸润的关系。方法肿瘤基因组图谱(the cancer genome at...目的通过生物信息学分析核受体亚家族6 A组成员1(nuclear receptor subfamily 6 group A member 1,NR6A1)在胃癌中的作用,旨在阐明NR6A1在胃癌中诊断和预后价值,也包括其与免疫细胞浸润的关系。方法肿瘤基因组图谱(the cancer genome atlas,TCGA)数据库用于分析NR6A1在胃癌组织与癌旁组织中的表达差异。利用Kaplan-Meier数据库分析NR6A1表达与胃癌患者生存预后的关系,使用R包建立诊断的受试者工作特征(receiver operating characteristic curve,ROC)曲线和列线图(nomogram)模型。cBioPortal和MethSurv数据库用于分析NR6A1基因突变和DNA甲基化,以及对胃癌患者生存预后的影响。仙桃学术用于分析NR6A1表达与免疫细胞浸润丰度的关系。STRING数据库分析与NR6A1蛋白质相互作用网络信息。基因本体论(gene ontology,GO)和京都基因和基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)富集分析NR6A1相关基因的功能。使用JASPAR和GRNdb数据库预测基因NR6A1上游的转录因子。NR6A1具有转录因子的功能,通过GRNdb数据库预测其下游靶基因,并富集分析相关靶基因的功能。结果TCGA数据库结果表明,NR6A1在胃癌中高表达;Kaplan-Meier数据库提示高表达NR6A1的胃癌患者预后差。ROC曲线分析表明,NR6A1可作为胃癌组织的诊断标志物(AUC=0.851)。基因突变分析,NR6A1基因突变率为2.70%,且发生基因突变的患者预后差。甲基化分析表明,在13个DNA甲基化CpG位点中,cg13409170的低甲基化与患者预后不良有关。免疫浸润分析表明,NR6A1高表达与多种免疫细胞浸润呈负相关。GO和KEGG分析主要富集在RNA转运和降解通路中。利用JASPAR和GRNdb数据库预测基因NR6A1的上游转录因子为KLF5和E2F6。GRNdb数据库预测出转录因子NR6A1的7个下游靶基因(PLEKHG5、GTF2IRD1、CEL、SPRY2、MSL1、HNF4A-AS1、ADNP-AS1),靶基因富集分析功能主要包括突触前易化、Ras蛋白信号转导调控、小GTP酶介导的信号导调控等通路。结论NR6A1在胃癌组织中高表达,其高表达、基因突变、低甲基化与患者预后差有关,且NR6A1和免疫细胞浸润丰度相关,可作为胃癌预后可靠的标志物。展开更多
核受体在细胞稳态的维持以及疾病的发生发展等方面发挥着重要的作用。为了探究核受体亚家族6A组成员1(nuclear receptor subfamily 6 group A member 1,NR6A1)在肝癌中的作用及机制,首先,分析了癌症基因组图谱(The Cancer Genome Atlas,...核受体在细胞稳态的维持以及疾病的发生发展等方面发挥着重要的作用。为了探究核受体亚家族6A组成员1(nuclear receptor subfamily 6 group A member 1,NR6A1)在肝癌中的作用及机制,首先,分析了癌症基因组图谱(The Cancer Genome Atlas,TCGA)等数据库信息,发现NR6A1在肝癌中异常高表达且与患者预后不良有关;然后,通过CCK-8(Cell Counting Kit-8)、5-乙炔基-2′-脱氧尿苷(5-ethynyl-2′-deoxyuridine,EdU)、划痕实验发现,干扰NR6A1使肝癌细胞的增殖明显受到抑制但不影响细胞的迁移;其次,通过免疫共沉淀、免疫荧光、RNA干扰和过表达等实验,鉴定出N-myc下游调控基因1(N-myc downstream-regulated gene 1,NDRG1)是NR6A1在肝癌中的互作蛋白质,二者在肝癌中的表达呈正相关,且NR6A1正调控NDRG1的表达;最后,利用功能拯救实验证实,干扰NDRG1可以抑制NR6A1过表达造成的肝癌细胞增殖增强的现象。综上可知,NR6A1通过与NDRG1相互结合且上调NDRG1的表达来发挥促癌作用。展开更多
为探索椎骨发育相关(Vertebrae Development Associated,VRTN)基因和核受体亚家族6A组成员1(Nuclear Receptor Subfamily 6 Group A Member 1,NR6A1)基因多态性与北京黑猪群体脊椎数的关联性,筛选脊椎数相关的重要分子标记,本研究利用PC...为探索椎骨发育相关(Vertebrae Development Associated,VRTN)基因和核受体亚家族6A组成员1(Nuclear Receptor Subfamily 6 Group A Member 1,NR6A1)基因多态性与北京黑猪群体脊椎数的关联性,筛选脊椎数相关的重要分子标记,本研究利用PCR技术和测序技术检测北京黑猪与胸椎数相关的重要候选基因VRTN和NR6A1的单核苷酸多态性(SNPs)位点,开展群体遗传学分析,并与胸椎数和腰椎数进行关联分析。结果显示:VRTN g.20311;0312ins291突变位点在北京黑猪群体中表现为ins/ins、ins/-、-/-3种基因型;g.19034A>C突变位点在北京黑猪群体中表现为AA、AC和CC3种基因型;NR6A1c.748C-T突变位点在北京黑猪群体中仅表现为TT基因型;关联分析结果表明,VRTN g.20311;0312ins291位点与北京黑猪的胸椎数和腰椎数无显著相关;g.19034A>C位点在北京黑猪中野生型个体(AA)的胸椎数显著低于突变纯合个体(CC),突变杂合个体(AC)的胸椎数与野生型个体和突变纯合个体差异均不显著;野生型个体(AA)的腰椎数显著高于突变纯合个体(CC),突变杂合个体(AC)的腰椎数与野生型个体和突变纯合个体的腰椎数差异均不显著。综上,VRTN基因g.19034A>C位点可作为潜在的北京黑猪多胸椎数分子标记。展开更多
旨在探讨生殖细胞核因子(nuclear receptor subfamily 6 group A member 1,NR6A1)和椎体发育同源物(vertebrae development homolog, VRTN)基因的多态性对苏姜猪体重、体长、体高、胸围、胸宽、臀宽和背膘厚7个生产性状的影响。试验利用...旨在探讨生殖细胞核因子(nuclear receptor subfamily 6 group A member 1,NR6A1)和椎体发育同源物(vertebrae development homolog, VRTN)基因的多态性对苏姜猪体重、体长、体高、胸围、胸宽、臀宽和背膘厚7个生产性状的影响。试验利用PCR扩增和酶切片段多态性技术检测苏姜猪及其亲本猪品种中NR6A1和VRTN基因的多态性,并将NR6A1和VRTN基因的多态性与苏姜猪的生产性状进行关联性分析。遗传多态性分析结果表明,NR6A1基因p.Pro192Leu (g.299084751 C>T)位点在姜曲海猪、枫泾猪和杜洛克猪群体中已经纯合,分别为CC、CC和TT基因型;在苏姜猪群体中有TT、TC和CC 3种基因型,基因型频率分别为0.397、0.482和0.121,基因型分布频率符合哈迪温伯格平衡(Hardy-Weinberg equilibrium, HWE,P>0.05),处于中度多态(0.250.05),VRTN基因对苏姜猪胸围、胸宽和臀宽均有显著影响(P<0.05),对其他性状无显著影响(P>0.05)。合并基因型对苏姜猪体长、胸围和背膘厚均有显著影响(P<0.05),对其他性状无显著影响(P>0.05)。综上可知,NR6A1和VRTN基因对苏姜猪的生产性状具有一定影响,为后续应用NR6A1和VRTN基因辅助选育苏姜猪提供参考。展开更多
Background and Aims:Generally acceptable prognostic models for hepatocellular carcinoma(HCC)are not available.This study aimed to establish a prognostic model for HCC by identifying immune-related differentially expre...Background and Aims:Generally acceptable prognostic models for hepatocellular carcinoma(HCC)are not available.This study aimed to establish a prognostic model for HCC by identifying immune-related differentially expressed genes(IR-DEGs)and to investigate the potential role of NR6A1 in the progression of HCC.Methods:Bioinformatics analysis using The Cancer Genome Atlas and ImmPort databases was used to identify IR-DEGs.Lasso Cox regression and multivariate Cox regression analysis were used to establish a prognostic model of HCC.Kaplan-Meier analysis and the receiver operating characteristic(ROC)curves were used to evaluate the performance of the prognostic model,which was further verified in the International Cancer Genome Consortium(ICGC)database.Gene set enrichment analysis was used to explore the potential pathways of NR6A1.Cell counting kit 8,colony formation,wound healing,and Transwell migration assays using Huh7 cells,and tumor formation models in nude mice were conducted.Results:A prognostic model established based on ten identified IR-DEGs including HSPA4,FABP6,MAPT,NDRG1,APLN,IL17D,LHB,SPP1,GLP1R,and NR6A1,effectively predicted the prognosis of HCC patients,was confirmed by the ROC curves and verified in ICGC database.NR6A1 expression was significantly up-regulated in HCC patients,and NR6A1 was significantly associated with a low survival rate.Gene set enrichment analysis showed the enrichment of cell cycle,mTOR,WNT,and ERBB signaling pathways in patients with high NR6A1 expression.NR6A1 promoted cell proliferation,invasiveness,migration,and malignant tumor formation and growth in vitro and in vivo.Conclusions:An effective prognostic model for HCC,based on a novel signature of 10 immune-related genes,was established.NR6A1 was up-regulated in HCC and was associated with a poor prognosis of HCC.NR6A1 promoted cell proliferation,migration,and growth of HCC,most likely through the cell cycle,mTOR,WNT,and ERBB signaling pathways.展开更多
Aim:Resistance to hormonal and targeted therapies in breast cancer limits treatment efficacy.Epigenetic alterations,including changes mediated by DNA methyltransferases,play a key role in this process.Previously,we id...Aim:Resistance to hormonal and targeted therapies in breast cancer limits treatment efficacy.Epigenetic alterations,including changes mediated by DNA methyltransferases,play a key role in this process.Previously,we identified that resistance to tamoxifen and rapamycin is associated with the suppression of DNMT3A.This study aims to further explore the mechanisms underlying this suppression,with a focus on identifying NR6A1 as a novel regulatory factor.Methods:Acquisition of resistant breast cancer cell sublines,MTT-test,immunoblotting,transient transfection and reporter analysis,lentiviral infection,qRT-PCR,and analysis of methylation using bisulfite pyrosequencing.Results:Our findings indicate that the development of cross-resistance in breast cancer cells to hormonal and targeted therapies involves a shift in cell signaling to alternative AKT pathways,marked by a localized suppression of the NR6A1/DNMT3A axis and associated DNA methylation changes.We demonstrated the critical role of NR6A1 downregulation in resistance development.Additionally,we observed activation of Snail-a key regulator in the epithelial-mesenchymal transition-as a mediator of the effects of NR6A1 depletion,establishing a direct link between Snail expression and resistance formation.Conclusion:The coordinated suppression of NR6A1 and DNMT3A may contribute to sustaining the resistant phenotype in breast cancer cells.This pathway could serve as a predictive marker,helping guide the selection of optimal therapeutic strategies for breast cancer treatment in the future.展开更多
文摘目的通过生物信息学分析核受体亚家族6 A组成员1(nuclear receptor subfamily 6 group A member 1,NR6A1)在胃癌中的作用,旨在阐明NR6A1在胃癌中诊断和预后价值,也包括其与免疫细胞浸润的关系。方法肿瘤基因组图谱(the cancer genome atlas,TCGA)数据库用于分析NR6A1在胃癌组织与癌旁组织中的表达差异。利用Kaplan-Meier数据库分析NR6A1表达与胃癌患者生存预后的关系,使用R包建立诊断的受试者工作特征(receiver operating characteristic curve,ROC)曲线和列线图(nomogram)模型。cBioPortal和MethSurv数据库用于分析NR6A1基因突变和DNA甲基化,以及对胃癌患者生存预后的影响。仙桃学术用于分析NR6A1表达与免疫细胞浸润丰度的关系。STRING数据库分析与NR6A1蛋白质相互作用网络信息。基因本体论(gene ontology,GO)和京都基因和基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)富集分析NR6A1相关基因的功能。使用JASPAR和GRNdb数据库预测基因NR6A1上游的转录因子。NR6A1具有转录因子的功能,通过GRNdb数据库预测其下游靶基因,并富集分析相关靶基因的功能。结果TCGA数据库结果表明,NR6A1在胃癌中高表达;Kaplan-Meier数据库提示高表达NR6A1的胃癌患者预后差。ROC曲线分析表明,NR6A1可作为胃癌组织的诊断标志物(AUC=0.851)。基因突变分析,NR6A1基因突变率为2.70%,且发生基因突变的患者预后差。甲基化分析表明,在13个DNA甲基化CpG位点中,cg13409170的低甲基化与患者预后不良有关。免疫浸润分析表明,NR6A1高表达与多种免疫细胞浸润呈负相关。GO和KEGG分析主要富集在RNA转运和降解通路中。利用JASPAR和GRNdb数据库预测基因NR6A1的上游转录因子为KLF5和E2F6。GRNdb数据库预测出转录因子NR6A1的7个下游靶基因(PLEKHG5、GTF2IRD1、CEL、SPRY2、MSL1、HNF4A-AS1、ADNP-AS1),靶基因富集分析功能主要包括突触前易化、Ras蛋白信号转导调控、小GTP酶介导的信号导调控等通路。结论NR6A1在胃癌组织中高表达,其高表达、基因突变、低甲基化与患者预后差有关,且NR6A1和免疫细胞浸润丰度相关,可作为胃癌预后可靠的标志物。
文摘核受体在细胞稳态的维持以及疾病的发生发展等方面发挥着重要的作用。为了探究核受体亚家族6A组成员1(nuclear receptor subfamily 6 group A member 1,NR6A1)在肝癌中的作用及机制,首先,分析了癌症基因组图谱(The Cancer Genome Atlas,TCGA)等数据库信息,发现NR6A1在肝癌中异常高表达且与患者预后不良有关;然后,通过CCK-8(Cell Counting Kit-8)、5-乙炔基-2′-脱氧尿苷(5-ethynyl-2′-deoxyuridine,EdU)、划痕实验发现,干扰NR6A1使肝癌细胞的增殖明显受到抑制但不影响细胞的迁移;其次,通过免疫共沉淀、免疫荧光、RNA干扰和过表达等实验,鉴定出N-myc下游调控基因1(N-myc downstream-regulated gene 1,NDRG1)是NR6A1在肝癌中的互作蛋白质,二者在肝癌中的表达呈正相关,且NR6A1正调控NDRG1的表达;最后,利用功能拯救实验证实,干扰NDRG1可以抑制NR6A1过表达造成的肝癌细胞增殖增强的现象。综上可知,NR6A1通过与NDRG1相互结合且上调NDRG1的表达来发挥促癌作用。
文摘为探索椎骨发育相关(Vertebrae Development Associated,VRTN)基因和核受体亚家族6A组成员1(Nuclear Receptor Subfamily 6 Group A Member 1,NR6A1)基因多态性与北京黑猪群体脊椎数的关联性,筛选脊椎数相关的重要分子标记,本研究利用PCR技术和测序技术检测北京黑猪与胸椎数相关的重要候选基因VRTN和NR6A1的单核苷酸多态性(SNPs)位点,开展群体遗传学分析,并与胸椎数和腰椎数进行关联分析。结果显示:VRTN g.20311;0312ins291突变位点在北京黑猪群体中表现为ins/ins、ins/-、-/-3种基因型;g.19034A>C突变位点在北京黑猪群体中表现为AA、AC和CC3种基因型;NR6A1c.748C-T突变位点在北京黑猪群体中仅表现为TT基因型;关联分析结果表明,VRTN g.20311;0312ins291位点与北京黑猪的胸椎数和腰椎数无显著相关;g.19034A>C位点在北京黑猪中野生型个体(AA)的胸椎数显著低于突变纯合个体(CC),突变杂合个体(AC)的胸椎数与野生型个体和突变纯合个体差异均不显著;野生型个体(AA)的腰椎数显著高于突变纯合个体(CC),突变杂合个体(AC)的腰椎数与野生型个体和突变纯合个体的腰椎数差异均不显著。综上,VRTN基因g.19034A>C位点可作为潜在的北京黑猪多胸椎数分子标记。
文摘旨在探讨生殖细胞核因子(nuclear receptor subfamily 6 group A member 1,NR6A1)和椎体发育同源物(vertebrae development homolog, VRTN)基因的多态性对苏姜猪体重、体长、体高、胸围、胸宽、臀宽和背膘厚7个生产性状的影响。试验利用PCR扩增和酶切片段多态性技术检测苏姜猪及其亲本猪品种中NR6A1和VRTN基因的多态性,并将NR6A1和VRTN基因的多态性与苏姜猪的生产性状进行关联性分析。遗传多态性分析结果表明,NR6A1基因p.Pro192Leu (g.299084751 C>T)位点在姜曲海猪、枫泾猪和杜洛克猪群体中已经纯合,分别为CC、CC和TT基因型;在苏姜猪群体中有TT、TC和CC 3种基因型,基因型频率分别为0.397、0.482和0.121,基因型分布频率符合哈迪温伯格平衡(Hardy-Weinberg equilibrium, HWE,P>0.05),处于中度多态(0.250.05),VRTN基因对苏姜猪胸围、胸宽和臀宽均有显著影响(P<0.05),对其他性状无显著影响(P>0.05)。合并基因型对苏姜猪体长、胸围和背膘厚均有显著影响(P<0.05),对其他性状无显著影响(P>0.05)。综上可知,NR6A1和VRTN基因对苏姜猪的生产性状具有一定影响,为后续应用NR6A1和VRTN基因辅助选育苏姜猪提供参考。
文摘Background and Aims:Generally acceptable prognostic models for hepatocellular carcinoma(HCC)are not available.This study aimed to establish a prognostic model for HCC by identifying immune-related differentially expressed genes(IR-DEGs)and to investigate the potential role of NR6A1 in the progression of HCC.Methods:Bioinformatics analysis using The Cancer Genome Atlas and ImmPort databases was used to identify IR-DEGs.Lasso Cox regression and multivariate Cox regression analysis were used to establish a prognostic model of HCC.Kaplan-Meier analysis and the receiver operating characteristic(ROC)curves were used to evaluate the performance of the prognostic model,which was further verified in the International Cancer Genome Consortium(ICGC)database.Gene set enrichment analysis was used to explore the potential pathways of NR6A1.Cell counting kit 8,colony formation,wound healing,and Transwell migration assays using Huh7 cells,and tumor formation models in nude mice were conducted.Results:A prognostic model established based on ten identified IR-DEGs including HSPA4,FABP6,MAPT,NDRG1,APLN,IL17D,LHB,SPP1,GLP1R,and NR6A1,effectively predicted the prognosis of HCC patients,was confirmed by the ROC curves and verified in ICGC database.NR6A1 expression was significantly up-regulated in HCC patients,and NR6A1 was significantly associated with a low survival rate.Gene set enrichment analysis showed the enrichment of cell cycle,mTOR,WNT,and ERBB signaling pathways in patients with high NR6A1 expression.NR6A1 promoted cell proliferation,invasiveness,migration,and malignant tumor formation and growth in vitro and in vivo.Conclusions:An effective prognostic model for HCC,based on a novel signature of 10 immune-related genes,was established.NR6A1 was up-regulated in HCC and was associated with a poor prognosis of HCC.NR6A1 promoted cell proliferation,migration,and growth of HCC,most likely through the cell cycle,mTOR,WNT,and ERBB signaling pathways.
基金supported by the Russian Science Foundation(project No.24-15-00173,https://rscf.ru/project/24-15-00173/).
文摘Aim:Resistance to hormonal and targeted therapies in breast cancer limits treatment efficacy.Epigenetic alterations,including changes mediated by DNA methyltransferases,play a key role in this process.Previously,we identified that resistance to tamoxifen and rapamycin is associated with the suppression of DNMT3A.This study aims to further explore the mechanisms underlying this suppression,with a focus on identifying NR6A1 as a novel regulatory factor.Methods:Acquisition of resistant breast cancer cell sublines,MTT-test,immunoblotting,transient transfection and reporter analysis,lentiviral infection,qRT-PCR,and analysis of methylation using bisulfite pyrosequencing.Results:Our findings indicate that the development of cross-resistance in breast cancer cells to hormonal and targeted therapies involves a shift in cell signaling to alternative AKT pathways,marked by a localized suppression of the NR6A1/DNMT3A axis and associated DNA methylation changes.We demonstrated the critical role of NR6A1 downregulation in resistance development.Additionally,we observed activation of Snail-a key regulator in the epithelial-mesenchymal transition-as a mediator of the effects of NR6A1 depletion,establishing a direct link between Snail expression and resistance formation.Conclusion:The coordinated suppression of NR6A1 and DNMT3A may contribute to sustaining the resistant phenotype in breast cancer cells.This pathway could serve as a predictive marker,helping guide the selection of optimal therapeutic strategies for breast cancer treatment in the future.
