BACKGROUND N6-methyladenosine(m6A)methylation modification exists in Epstein-Barr virus(EBV)primary infection,latency,and lytic reactivation.It also modifies EBV latent genes and lytic genes.EBV-associated gastric can...BACKGROUND N6-methyladenosine(m6A)methylation modification exists in Epstein-Barr virus(EBV)primary infection,latency,and lytic reactivation.It also modifies EBV latent genes and lytic genes.EBV-associated gastric cancer(EBVaGC)is a distinctive molecular subtype of GC.We hypothesized EBV and m6A methylation regulators interact with each other in EBVaGC to differentiate it from other types of GC.AIM To investigate the mechanisms of m6A methylation regulators in EBVaGC to determine the differentiating factors from other types of GC.METHODS First,The Cancer Gene Atlas and Gene Expression Omnibus databases were used to analyze the expression pattern of m6A methylation regulators between EBVaGC and EBV-negative GC(EBVnGC).Second,we identified Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment of m6A-related differentially expressed genes.We quantified the relative abundance of immune cells and inflammatory factors in the tumor microenvironment(TME).Finally,cell counting kit-8 cell proliferation test,transwell test,and flow cytometry were used to verify the effect of insulin-like growth factor binding protein 1(IGFBP1)in EBVaGC cell lines.RESULTS m6A methylation regulators were involved in the occurrence and development of EBVaGC.Compared with EBVnGC,the expression levels of m6A methylation regulators Wilms tumor 1-associated protein,RNA binding motif protein 15B,CBL proto-oncogene like 1,leucine rich pentatricopeptide repeat containing,heterogeneous nuclear ribonucleoprotein A2B1,IGFBP1,and insulin-like growth factor 2 binding protein 1 were significantly downregulated in EBVaGC(P<0.05).The overall survival rate of EBVaGC patients with a lower expression level of IGFBP1 was significantly higher(P=0.046).GO and KEGG functional enrichment analyses showed that the immunity pathways were significantly activated and rich in immune cell infiltration in EBVaGC.Compared with EBVnGC,the infiltration of activated CD4+T cells,activated CD8+T cells,monocytes,activated dendritic cells,and plasmacytoid dendritic cells were significantly upregulated in EBVaGC(P<0.001).In EBVaGC,the expression level of proinflammatory factors interleukin(IL)-17,IL-21,and interferon-γ and immunosuppressive factor IL-10 were significantly increased(P<0.05).In vitro experiments demonstrated that the expression level of IGFBP1 was significantly lower in an EBVaGC cell line(SNU719)than in an EBVnGC cell line(AGS)(P<0.05).IGFBP1 overexpression significantly attenuated proliferation and migration and promoted the apoptosis levels in SNU719.Interfering IGFBP1 significantly promoted proliferation and migration and attenuated the apoptosis levels in AGS.CONCLUSION m6A regulators could remodel the TME of EBVaGC,which is classified as an immune-inflamed phenotype and referred to as a“hot”tumor.Among these regulators,we demonstrated that IGFBP1 affected proliferation,migration,and apoptosis.展开更多
目的探索伴有焦虑症状双相抑郁患者认知功能与N-甲基-D-天冬氨酸受体2B亚基(glutamate iono-tropic receptor NMDA type subunit 2B,GRIN2B)基因启动子区各CpG位点甲基化水平的相关性。方法根据汉密尔顿焦虑量表(14-item Hamilton anxie...目的探索伴有焦虑症状双相抑郁患者认知功能与N-甲基-D-天冬氨酸受体2B亚基(glutamate iono-tropic receptor NMDA type subunit 2B,GRIN2B)基因启动子区各CpG位点甲基化水平的相关性。方法根据汉密尔顿焦虑量表(14-item Hamilton anxiety rating scale,HAMA)评分将31例双相抑郁患者分为焦虑组15例和非焦虑组16例,同期选取16名健康对照。采用蒙特利尔认知评估量表(Montreal cognitive assessment,MoCA)、数值广度测验(digital span test,DST)、连线测试A部分(trail making test A,TMT-A)、斯特鲁普色词测验(Stroop color and word test,SCWT)评估3组总体认知功能、注意力及执行控制、信息处理速度、执行功能等认知功能维度,采用Massarray质谱法检测所有受试者外周血GRIN2B基因启动子区各CpG位点的DNA甲基化水平。结果3组GRIN2B基因启动子区DNA甲基化水平差异性位点为CpG3、CpG5、CpG7、CpG10、CpG12(P<0.05),其中,焦虑组CpG12甲基化水平低于非焦虑组(36.23%±16.41%vs.50.20%±19.79%,P<0.05)。偏相关分析显示,焦虑组患者中较差的命名能力与GRIN2B基因CpG4低甲基化水平相关(r=0.670,P=0.034),较差的执行功能与CpG6低甲基化水平相关(r=0.926,P<0.001),较差的注意力与GRIN2B基因CpG8高甲基化水平相关(r=-0.810,P=0.025),较差的言语记忆与CpG9高甲基化水平相关(r=-0.810,P<0.001),较差的抽象能力与CpG10高甲基化水平相关(r=-0.756,P=0.011)。结论GRIN2B基因启动子区DNA甲基化水平与伴有焦虑症状双相抑郁患者认知功能损害可能有关联,与双相抑郁患者焦虑症状的产生也可能有关联。展开更多
基金Supported by the Sub-Project of the National Key Research and Development Program,No.2021YFC2600263.
文摘BACKGROUND N6-methyladenosine(m6A)methylation modification exists in Epstein-Barr virus(EBV)primary infection,latency,and lytic reactivation.It also modifies EBV latent genes and lytic genes.EBV-associated gastric cancer(EBVaGC)is a distinctive molecular subtype of GC.We hypothesized EBV and m6A methylation regulators interact with each other in EBVaGC to differentiate it from other types of GC.AIM To investigate the mechanisms of m6A methylation regulators in EBVaGC to determine the differentiating factors from other types of GC.METHODS First,The Cancer Gene Atlas and Gene Expression Omnibus databases were used to analyze the expression pattern of m6A methylation regulators between EBVaGC and EBV-negative GC(EBVnGC).Second,we identified Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)functional enrichment of m6A-related differentially expressed genes.We quantified the relative abundance of immune cells and inflammatory factors in the tumor microenvironment(TME).Finally,cell counting kit-8 cell proliferation test,transwell test,and flow cytometry were used to verify the effect of insulin-like growth factor binding protein 1(IGFBP1)in EBVaGC cell lines.RESULTS m6A methylation regulators were involved in the occurrence and development of EBVaGC.Compared with EBVnGC,the expression levels of m6A methylation regulators Wilms tumor 1-associated protein,RNA binding motif protein 15B,CBL proto-oncogene like 1,leucine rich pentatricopeptide repeat containing,heterogeneous nuclear ribonucleoprotein A2B1,IGFBP1,and insulin-like growth factor 2 binding protein 1 were significantly downregulated in EBVaGC(P<0.05).The overall survival rate of EBVaGC patients with a lower expression level of IGFBP1 was significantly higher(P=0.046).GO and KEGG functional enrichment analyses showed that the immunity pathways were significantly activated and rich in immune cell infiltration in EBVaGC.Compared with EBVnGC,the infiltration of activated CD4+T cells,activated CD8+T cells,monocytes,activated dendritic cells,and plasmacytoid dendritic cells were significantly upregulated in EBVaGC(P<0.001).In EBVaGC,the expression level of proinflammatory factors interleukin(IL)-17,IL-21,and interferon-γ and immunosuppressive factor IL-10 were significantly increased(P<0.05).In vitro experiments demonstrated that the expression level of IGFBP1 was significantly lower in an EBVaGC cell line(SNU719)than in an EBVnGC cell line(AGS)(P<0.05).IGFBP1 overexpression significantly attenuated proliferation and migration and promoted the apoptosis levels in SNU719.Interfering IGFBP1 significantly promoted proliferation and migration and attenuated the apoptosis levels in AGS.CONCLUSION m6A regulators could remodel the TME of EBVaGC,which is classified as an immune-inflamed phenotype and referred to as a“hot”tumor.Among these regulators,we demonstrated that IGFBP1 affected proliferation,migration,and apoptosis.
文摘目的探索伴有焦虑症状双相抑郁患者认知功能与N-甲基-D-天冬氨酸受体2B亚基(glutamate iono-tropic receptor NMDA type subunit 2B,GRIN2B)基因启动子区各CpG位点甲基化水平的相关性。方法根据汉密尔顿焦虑量表(14-item Hamilton anxiety rating scale,HAMA)评分将31例双相抑郁患者分为焦虑组15例和非焦虑组16例,同期选取16名健康对照。采用蒙特利尔认知评估量表(Montreal cognitive assessment,MoCA)、数值广度测验(digital span test,DST)、连线测试A部分(trail making test A,TMT-A)、斯特鲁普色词测验(Stroop color and word test,SCWT)评估3组总体认知功能、注意力及执行控制、信息处理速度、执行功能等认知功能维度,采用Massarray质谱法检测所有受试者外周血GRIN2B基因启动子区各CpG位点的DNA甲基化水平。结果3组GRIN2B基因启动子区DNA甲基化水平差异性位点为CpG3、CpG5、CpG7、CpG10、CpG12(P<0.05),其中,焦虑组CpG12甲基化水平低于非焦虑组(36.23%±16.41%vs.50.20%±19.79%,P<0.05)。偏相关分析显示,焦虑组患者中较差的命名能力与GRIN2B基因CpG4低甲基化水平相关(r=0.670,P=0.034),较差的执行功能与CpG6低甲基化水平相关(r=0.926,P<0.001),较差的注意力与GRIN2B基因CpG8高甲基化水平相关(r=-0.810,P=0.025),较差的言语记忆与CpG9高甲基化水平相关(r=-0.810,P<0.001),较差的抽象能力与CpG10高甲基化水平相关(r=-0.756,P=0.011)。结论GRIN2B基因启动子区DNA甲基化水平与伴有焦虑症状双相抑郁患者认知功能损害可能有关联,与双相抑郁患者焦虑症状的产生也可能有关联。
