YT521-B同源结构域家族成员2(YTH domain family protein2,YTHDF2)是一种m6A阅读蛋白,可通过与m6A修饰的RNA特异性结合并介导其降解,从而影响基因表达,参与多种生理和病理过程。目前发现YTHDF2在炎性反应和多种炎症性疾病中的异常表达,...YT521-B同源结构域家族成员2(YTH domain family protein2,YTHDF2)是一种m6A阅读蛋白,可通过与m6A修饰的RNA特异性结合并介导其降解,从而影响基因表达,参与多种生理和病理过程。目前发现YTHDF2在炎性反应和多种炎症性疾病中的异常表达,通过调节靶RNA的表达和信号通路的激活参与炎症性疾病的发生、发展。本文就YTHDF2的结构、功能以及在炎症性疾病发生、发展与药物治疗效应中的作用和相关分子发生机制进行综述。展开更多
目的研究敲低YTH结构域N^6甲基腺嘌呤(m^6A)RNA结合蛋白2(YTHDF2)对MGC-803人胃癌细胞的增殖、细胞周期和细胞凋亡的影响。方法用UCSC Cancer Browser下载癌症基因组图谱(TCGA)数据库筛查YTHDF2 mRNA在胃癌中的表达情况。设计并构建能...目的研究敲低YTH结构域N^6甲基腺嘌呤(m^6A)RNA结合蛋白2(YTHDF2)对MGC-803人胃癌细胞的增殖、细胞周期和细胞凋亡的影响。方法用UCSC Cancer Browser下载癌症基因组图谱(TCGA)数据库筛查YTHDF2 mRNA在胃癌中的表达情况。设计并构建能够靶向敲低YTHDF2的短发夹RNA(shRNA)病毒载体;病毒感染MGC-803细胞,敲低YTHDF2水平后,实时荧光定量PCR检测YTHDF2 mRNA水平,Western blot法检测YTHDF2蛋白水平,CCK-8法检测细胞增殖,流式细胞术检测细胞周期和细胞凋亡。结果通过TCGA数据库查询发现YTHDF2在胃癌组织中高表达。成功构建YTHDF2敲低的稳定转染MGC-803细胞。与对照组相比,YTHDF2敲低后,明显抑制细胞增殖;G1期细胞明显增加,S期细胞明显减少;细胞凋亡率升高。结论敲低MGC-803胃癌细胞YTHDF2水平,抑制胃癌细胞增殖并促进其凋亡。展开更多
目的探讨新型RNA甲基化修饰(N6-methyladenosine, m6A)阅读器蛋白YTH结构域m6A结合蛋白2(YTHDF2)在卵巢癌(Ovarian cancer)的表达水平及增殖调控作用。方法利用人类蛋白质图谱(The human protein atlas, HPA)分析YTHDF2在卵巢癌组织的...目的探讨新型RNA甲基化修饰(N6-methyladenosine, m6A)阅读器蛋白YTH结构域m6A结合蛋白2(YTHDF2)在卵巢癌(Ovarian cancer)的表达水平及增殖调控作用。方法利用人类蛋白质图谱(The human protein atlas, HPA)分析YTHDF2在卵巢癌组织的表达定位;收集卵巢癌和癌旁组织标本31对,应用实时定量PCR方法检测YTHDF2转录水平;CCK-8方法检测细胞增殖水平。结果 YTHDF2蛋白主要表达于肿瘤细胞核;与癌旁组织比较,YTHDF2在卵巢癌组织中的表达明显增加(P<0.05);转染YTHDF2 siRNA能够显著降低卵巢癌细胞YTHDF2的表达水平,抑制细胞增殖。结论 YTHDF2是一个新的卵巢癌生物标记物,可作为潜在的治疗新靶点。展开更多
目的探讨敲低YTH结构域N^6甲基腺嘌呤(m^6A)RNA结合蛋白2(YTHDF2)对人宫颈癌细胞增殖、周期和凋亡的影响。方法利用人蛋白图谱(Human Protein Atlas)数据库分析YTHDF2在宫颈癌中表达及与生存时间的关系。收集宫颈癌和正常宫颈组织各31例...目的探讨敲低YTH结构域N^6甲基腺嘌呤(m^6A)RNA结合蛋白2(YTHDF2)对人宫颈癌细胞增殖、周期和凋亡的影响。方法利用人蛋白图谱(Human Protein Atlas)数据库分析YTHDF2在宫颈癌中表达及与生存时间的关系。收集宫颈癌和正常宫颈组织各31例,采用免疫组织化学法检测YTHDF2的蛋白表达差异;利用敲低YTHDF2的短发夹RNA(shRNA)及空载质粒包装慢病毒,并感染至宫颈癌HeLa细胞和SiHa细胞,实时荧光定量PCR及Western blot法检测YTHDF2的mRNA及蛋白水平,敲低YTHDF2后,采用CCK-8法检测细胞增殖活性,集落形成实验检测细胞集落形成能力;流式细胞术检测细胞周期和细胞凋亡情况。结果检测数据库发现YTHDF2在宫颈癌中表达越高生存时间越短,与正常宫颈组织相比,YTHDF2在宫颈癌组织高表达。敲低YTHDF2后,抑制宫颈癌细胞增殖,促进细胞凋亡,使细胞阻滞于S期。结论YTHDF2在宫颈癌组织中高表达,敲低后抑制宫颈癌细胞增殖并促进其凋亡。展开更多
目的:探讨m6A甲基化修饰结合蛋白YTHDF2(YTH domain-containing family protein 2)在肝癌(Hepatocellular carcinoma,HCC)组织中的表达及临床意义。方法:1)采用Western blot和实时荧光定量PCR(Quantitative Real Time PCR, qRT-PCR)检...目的:探讨m6A甲基化修饰结合蛋白YTHDF2(YTH domain-containing family protein 2)在肝癌(Hepatocellular carcinoma,HCC)组织中的表达及临床意义。方法:1)采用Western blot和实时荧光定量PCR(Quantitative Real Time PCR, qRT-PCR)检测20对肝癌和癌旁组织中YTHDF2在蛋白和m RNA水平的表达情况。2)通过免疫组织化学(Immunochemistry, IHC)检测40对肝癌和癌旁组织芯片YTHDF2的表达情况,并以H-score评分法进行半定量分析。3)通过基因表达谱数据动态分析数据库GEPIA(http://gepia.cancer-pku.cn/)分析YTHDF2在肝癌组织中的表达及对患者生存预后的影响;通过肿瘤免疫评估资源数据库TIMER(https://cistrome.shinyapps.io/timer/)分析YTHDF2与肝癌免疫微环境的相关性。结果:相比于癌旁肝组织,YTHDF2在肝癌组织中蛋白和m RNA水平均显著高表达(P<0.05);GEPIA数据库分析验证:YTHDF2肝癌组织表达水平高于正常肝组织,且YTHDF2在肝癌不同分期中表达水平均高于正常肝组织(P=0.0206),Stage Ⅲ期最为明显;YTHDF2高表达患者的总体生存率(Overall Survival,OS)和无复发生存率(Disease Free Survival, DFS)均较差(P=0.00027和P=0.013);TIMER数据库分析表明:YTHDF2在肝癌免疫微环境中与各类免疫细胞呈正相关(P<0.05),但肝癌患者的累积存活率不受免疫细胞的影响(P>0.05),YTHDF2高表达对患者的生存预后具有不良影响(P=0.007)。结论:YTHDF2在肝癌组织中显著高表达,且YTHDF2高表达水平对肝癌患者生存预后具有不良影响,YTHDF2可作为肝癌临床预后判断的分子标志物,为今后肝癌治疗靶点提供新的策略。展开更多
Atrial fibrillation(AF)is the most common arrhythmia,which is tightly associated with the abnormal expression and function of ion channels in the atrial cardiomyocytes.N^(6)-methyladenosine(m^(6)A),a widespread chemic...Atrial fibrillation(AF)is the most common arrhythmia,which is tightly associated with the abnormal expression and function of ion channels in the atrial cardiomyocytes.N^(6)-methyladenosine(m^(6)A),a widespread chemical modification in eukaryotic mRNA,is known to play a significant regulatory role in the pathogenesis of heart disease.However,the significance of m6A regulatory proteins in the onset of AF remains unclear.Here,we demonstrate that the m^(6)A reader protein YTHDF2 regulates atrial electrical remodeling and AF onset by modulating the Cav1.2 expression.Firstly,YTHDF2 expression was selectively upregulated in rat atrial cardiomyocytes with AF.Secondly,YTHDF2 knockout reduced AF susceptibility in mice.Thirdly,the knockout of YTHDF2 increased Cav1.2 protein levels in an m^(6)A-independent manner,ultimately prolonging the atrial myocardial refractory period,a critical electrophysiological substrate for the onset of AF.Fourthly,the N-terminal domain of YTHDF2 was identified as critical for Cacna1c mRNA translation regulation.Overall,our findings unveil that YTHDF2 can alter Cav1.2 protein expression in an m^(6)A-independent manner,thereby facilitating the onset of AF.Our study suggests that YTHDF2 may be a potential intervention target for AF.展开更多
N6-methyladenosine(m^(6)A)plays crucial roles in development and cellular reprogramming.During embryonic development,pluripotency transitions from a naïve to a primed state,and modeling the reverse primed-to-na&#...N6-methyladenosine(m^(6)A)plays crucial roles in development and cellular reprogramming.During embryonic development,pluripotency transitions from a naïve to a primed state,and modeling the reverse primed-to-naïve transition(PNT)provides a valuable framework for investigating pluripotency regulation.Here,we show that inhibiting METTL3 significantly promotes PNT in an m^(6)A-dependent manner.Mechanistically,we found that suppressing METTL3 and YTHDF2 prolongs the lifetimes of pluripotency-associated mRNAs,such as Nanog and Sox2,during PNT.In addition,Gstp1 was identified as a downstream target of METTL3 inhibition and YTHDF2 knockout.Gstp1 overexpression enhances PNT,whereas its inhibition impedes the transition.Overall,our findings suggest that YTHDF2 facilitates the removal of pluripotency gene transcripts and Gstp1,thereby promoting PNT reprogramming through m^(6)A-mediated posttranscriptional control.展开更多
目的:利用生物信息学和免疫组化方法分析YTHDF1和YTHDF2在肺鳞癌(lung squamous cell carcinoma,LUSC)组织中的表达水平及临床意义。方法:利用TCGA数据库对LUSC中YTHDF1和YTHDF2的表达和相关性进行分析。通过基因集富集分析(gene set en...目的:利用生物信息学和免疫组化方法分析YTHDF1和YTHDF2在肺鳞癌(lung squamous cell carcinoma,LUSC)组织中的表达水平及临床意义。方法:利用TCGA数据库对LUSC中YTHDF1和YTHDF2的表达和相关性进行分析。通过基因集富集分析(gene set enrichment analysis,GSEA)揭示YTHDF1和YTHDF2可能涉及的生物学功能和信号通路。最后采用免疫组化方法证实YTHDF1和YTHDF2在LUSC组织样本中的表达,探讨YTHDF1和YTHDF2表达水平与病理特征的关系,并用统计学方法分析其与PD-L1的相关性。结果:(1)YTHDF1和YTHDF2在LUSC组织中均高于癌旁组织(P<0.05);(2)YTHDF1与T分期、肿瘤分期存在相关性(P<0.05);(3)年龄是影响LUSC患者总生存期(over all survival,OS)的独立危险因素,YTHDF1表达量与T分期是影响疾病特异生存期(disease specific survival,DSS)的独立危险因素;(4)YTHDF1和YTHDF2高表达LUSC患者的基因富集主要在治疗靶点上通路上;在低表达YTHDF1和YTHDF2组中,基因富集在免疫活性相关方面。(5)YTHDF2与PD-L1表达呈正相关(P<0.05)。结论:YTHDF1和YTHDF2的表达水平可能影响LUSC免疫微环境,同时也提示了其可能是LUSC免疫治疗的潜在靶点和生物标志物。展开更多
As the most common internal modification of mRNA,Ne-methyladenosine(m^(6)A)and its regulators modulate gene expression and play critical roles in various biological and patholog-ical processes including tumorigenesis....As the most common internal modification of mRNA,Ne-methyladenosine(m^(6)A)and its regulators modulate gene expression and play critical roles in various biological and patholog-ical processes including tumorigenesis.It was reported previously that m^(6)A methyltransferase(writer),methyltransferase-like 3(METTL3)adds m^(6)A in primary microRNAs(pri-miRNAs)and fa-cilitates its processing into precursor miRNAs(pre-miRNAs).However,it is unknown whether m^(6)A modification also plays a role in the maturation process of pre-miRNAs and(if so)whether such a function contributes to tumorigenesis.Here,we found that YTHDF2 is aberrantly overexpressed in acute myeloid leukemia(AML)patients,especially in relapsed patients,and plays an onco-genic role in AML.Moreover,YTHDF2 promotes expression of miR-126-3p(also known as miR-126,as it is the main product of precursor miR-126(pre-miR-126)),a miRNA that was reported as an oncomiRNA in AML,through facilitating the processing of pre-miR-126 into mature miR-126.Mechanistically,YTHDF2 recognizes m^(6)A modification in pre-miR-126 and recruits AGO2,a regulator of pre-miRNA processing,to promote the maturation of pre-miR-126.YTHDF2 posi-tively and negatively correlates with miR-126 and miR-126's downstream target genes,respec-tively,in AML patients,and forced expression of miR-126 could largely rescue YTHDF2/Ythdf2 depletion-mediated suppression on AML cell growth/proliferation and leukemogenesis,indi-cating that miR-126 is a functionally important target of YTHDF2 in AML.Overall,our studies not only reveal a previously unappreciated YTHDF2/miR-126 axis in AML and highlight the ther-apeutic potential of targeting this axis for AML treatment,but also suggest that m^(6)A plays a role in pre-miRNA processing that contributes to tumorigenesis.展开更多
文摘YT521-B同源结构域家族成员2(YTH domain family protein2,YTHDF2)是一种m6A阅读蛋白,可通过与m6A修饰的RNA特异性结合并介导其降解,从而影响基因表达,参与多种生理和病理过程。目前发现YTHDF2在炎性反应和多种炎症性疾病中的异常表达,通过调节靶RNA的表达和信号通路的激活参与炎症性疾病的发生、发展。本文就YTHDF2的结构、功能以及在炎症性疾病发生、发展与药物治疗效应中的作用和相关分子发生机制进行综述。
文摘目的探讨新型RNA甲基化修饰(N6-methyladenosine, m6A)阅读器蛋白YTH结构域m6A结合蛋白2(YTHDF2)在卵巢癌(Ovarian cancer)的表达水平及增殖调控作用。方法利用人类蛋白质图谱(The human protein atlas, HPA)分析YTHDF2在卵巢癌组织的表达定位;收集卵巢癌和癌旁组织标本31对,应用实时定量PCR方法检测YTHDF2转录水平;CCK-8方法检测细胞增殖水平。结果 YTHDF2蛋白主要表达于肿瘤细胞核;与癌旁组织比较,YTHDF2在卵巢癌组织中的表达明显增加(P<0.05);转染YTHDF2 siRNA能够显著降低卵巢癌细胞YTHDF2的表达水平,抑制细胞增殖。结论 YTHDF2是一个新的卵巢癌生物标记物,可作为潜在的治疗新靶点。
文摘目的探讨敲低YTH结构域N^6甲基腺嘌呤(m^6A)RNA结合蛋白2(YTHDF2)对人宫颈癌细胞增殖、周期和凋亡的影响。方法利用人蛋白图谱(Human Protein Atlas)数据库分析YTHDF2在宫颈癌中表达及与生存时间的关系。收集宫颈癌和正常宫颈组织各31例,采用免疫组织化学法检测YTHDF2的蛋白表达差异;利用敲低YTHDF2的短发夹RNA(shRNA)及空载质粒包装慢病毒,并感染至宫颈癌HeLa细胞和SiHa细胞,实时荧光定量PCR及Western blot法检测YTHDF2的mRNA及蛋白水平,敲低YTHDF2后,采用CCK-8法检测细胞增殖活性,集落形成实验检测细胞集落形成能力;流式细胞术检测细胞周期和细胞凋亡情况。结果检测数据库发现YTHDF2在宫颈癌中表达越高生存时间越短,与正常宫颈组织相比,YTHDF2在宫颈癌组织高表达。敲低YTHDF2后,抑制宫颈癌细胞增殖,促进细胞凋亡,使细胞阻滞于S期。结论YTHDF2在宫颈癌组织中高表达,敲低后抑制宫颈癌细胞增殖并促进其凋亡。
文摘目的:探讨m6A甲基化修饰结合蛋白YTHDF2(YTH domain-containing family protein 2)在肝癌(Hepatocellular carcinoma,HCC)组织中的表达及临床意义。方法:1)采用Western blot和实时荧光定量PCR(Quantitative Real Time PCR, qRT-PCR)检测20对肝癌和癌旁组织中YTHDF2在蛋白和m RNA水平的表达情况。2)通过免疫组织化学(Immunochemistry, IHC)检测40对肝癌和癌旁组织芯片YTHDF2的表达情况,并以H-score评分法进行半定量分析。3)通过基因表达谱数据动态分析数据库GEPIA(http://gepia.cancer-pku.cn/)分析YTHDF2在肝癌组织中的表达及对患者生存预后的影响;通过肿瘤免疫评估资源数据库TIMER(https://cistrome.shinyapps.io/timer/)分析YTHDF2与肝癌免疫微环境的相关性。结果:相比于癌旁肝组织,YTHDF2在肝癌组织中蛋白和m RNA水平均显著高表达(P<0.05);GEPIA数据库分析验证:YTHDF2肝癌组织表达水平高于正常肝组织,且YTHDF2在肝癌不同分期中表达水平均高于正常肝组织(P=0.0206),Stage Ⅲ期最为明显;YTHDF2高表达患者的总体生存率(Overall Survival,OS)和无复发生存率(Disease Free Survival, DFS)均较差(P=0.00027和P=0.013);TIMER数据库分析表明:YTHDF2在肝癌免疫微环境中与各类免疫细胞呈正相关(P<0.05),但肝癌患者的累积存活率不受免疫细胞的影响(P>0.05),YTHDF2高表达对患者的生存预后具有不良影响(P=0.007)。结论:YTHDF2在肝癌组织中显著高表达,且YTHDF2高表达水平对肝癌患者生存预后具有不良影响,YTHDF2可作为肝癌临床预后判断的分子标志物,为今后肝癌治疗靶点提供新的策略。
基金supported by the National Natural Science Foundation of China(82121001,82222008,82070338,82470330)the Fundamental Research Funds for the Central Universities.
文摘Atrial fibrillation(AF)is the most common arrhythmia,which is tightly associated with the abnormal expression and function of ion channels in the atrial cardiomyocytes.N^(6)-methyladenosine(m^(6)A),a widespread chemical modification in eukaryotic mRNA,is known to play a significant regulatory role in the pathogenesis of heart disease.However,the significance of m6A regulatory proteins in the onset of AF remains unclear.Here,we demonstrate that the m^(6)A reader protein YTHDF2 regulates atrial electrical remodeling and AF onset by modulating the Cav1.2 expression.Firstly,YTHDF2 expression was selectively upregulated in rat atrial cardiomyocytes with AF.Secondly,YTHDF2 knockout reduced AF susceptibility in mice.Thirdly,the knockout of YTHDF2 increased Cav1.2 protein levels in an m^(6)A-independent manner,ultimately prolonging the atrial myocardial refractory period,a critical electrophysiological substrate for the onset of AF.Fourthly,the N-terminal domain of YTHDF2 was identified as critical for Cacna1c mRNA translation regulation.Overall,our findings unveil that YTHDF2 can alter Cav1.2 protein expression in an m^(6)A-independent manner,thereby facilitating the onset of AF.Our study suggests that YTHDF2 may be a potential intervention target for AF.
基金supported by the National Key R&D Program of China(2021YFA1102200,2024YFA1802300 and 2024YFA1107000)The National Natural Science Foundation of China(32225012)+4 种基金Major Project of Guangzhou National Laboratory(GZNL2023A02005)Guangdong Basic and Applied Basic Research Foundation(2025A1515012426,2023A1515010420)Science and Technology Projects in Guangzhou(2023A04J0724)Science and Technology Planning Project of Guangdong Province,China(2023B1212060050,2023B1212120009)Health@InnoHK Program launched by Innovation Technology Commission of the Hong Kong SAR,P.R.China.
文摘N6-methyladenosine(m^(6)A)plays crucial roles in development and cellular reprogramming.During embryonic development,pluripotency transitions from a naïve to a primed state,and modeling the reverse primed-to-naïve transition(PNT)provides a valuable framework for investigating pluripotency regulation.Here,we show that inhibiting METTL3 significantly promotes PNT in an m^(6)A-dependent manner.Mechanistically,we found that suppressing METTL3 and YTHDF2 prolongs the lifetimes of pluripotency-associated mRNAs,such as Nanog and Sox2,during PNT.In addition,Gstp1 was identified as a downstream target of METTL3 inhibition and YTHDF2 knockout.Gstp1 overexpression enhances PNT,whereas its inhibition impedes the transition.Overall,our findings suggest that YTHDF2 facilitates the removal of pluripotency gene transcripts and Gstp1,thereby promoting PNT reprogramming through m^(6)A-mediated posttranscriptional control.
文摘目的:利用生物信息学和免疫组化方法分析YTHDF1和YTHDF2在肺鳞癌(lung squamous cell carcinoma,LUSC)组织中的表达水平及临床意义。方法:利用TCGA数据库对LUSC中YTHDF1和YTHDF2的表达和相关性进行分析。通过基因集富集分析(gene set enrichment analysis,GSEA)揭示YTHDF1和YTHDF2可能涉及的生物学功能和信号通路。最后采用免疫组化方法证实YTHDF1和YTHDF2在LUSC组织样本中的表达,探讨YTHDF1和YTHDF2表达水平与病理特征的关系,并用统计学方法分析其与PD-L1的相关性。结果:(1)YTHDF1和YTHDF2在LUSC组织中均高于癌旁组织(P<0.05);(2)YTHDF1与T分期、肿瘤分期存在相关性(P<0.05);(3)年龄是影响LUSC患者总生存期(over all survival,OS)的独立危险因素,YTHDF1表达量与T分期是影响疾病特异生存期(disease specific survival,DSS)的独立危险因素;(4)YTHDF1和YTHDF2高表达LUSC患者的基因富集主要在治疗靶点上通路上;在低表达YTHDF1和YTHDF2组中,基因富集在免疫活性相关方面。(5)YTHDF2与PD-L1表达呈正相关(P<0.05)。结论:YTHDF1和YTHDF2的表达水平可能影响LUSC免疫微环境,同时也提示了其可能是LUSC免疫治疗的潜在靶点和生物标志物。
基金supported in part by the U.S.National Institutes of Health(NIH)grants R01 CA243386(J.C.),R01 CA271497,R01 CA214965(J.C.),R01 CA236399(J.C.),R01 DK124116(J.C)The Simms/Mann Family Foundation(J.C.).J.C.is a Leukemia&Lymphoma Society(LLS)Scholar and is supported by the LLS 2022 Scholar CDP Achievement Award.
文摘As the most common internal modification of mRNA,Ne-methyladenosine(m^(6)A)and its regulators modulate gene expression and play critical roles in various biological and patholog-ical processes including tumorigenesis.It was reported previously that m^(6)A methyltransferase(writer),methyltransferase-like 3(METTL3)adds m^(6)A in primary microRNAs(pri-miRNAs)and fa-cilitates its processing into precursor miRNAs(pre-miRNAs).However,it is unknown whether m^(6)A modification also plays a role in the maturation process of pre-miRNAs and(if so)whether such a function contributes to tumorigenesis.Here,we found that YTHDF2 is aberrantly overexpressed in acute myeloid leukemia(AML)patients,especially in relapsed patients,and plays an onco-genic role in AML.Moreover,YTHDF2 promotes expression of miR-126-3p(also known as miR-126,as it is the main product of precursor miR-126(pre-miR-126)),a miRNA that was reported as an oncomiRNA in AML,through facilitating the processing of pre-miR-126 into mature miR-126.Mechanistically,YTHDF2 recognizes m^(6)A modification in pre-miR-126 and recruits AGO2,a regulator of pre-miRNA processing,to promote the maturation of pre-miR-126.YTHDF2 posi-tively and negatively correlates with miR-126 and miR-126's downstream target genes,respec-tively,in AML patients,and forced expression of miR-126 could largely rescue YTHDF2/Ythdf2 depletion-mediated suppression on AML cell growth/proliferation and leukemogenesis,indi-cating that miR-126 is a functionally important target of YTHDF2 in AML.Overall,our studies not only reveal a previously unappreciated YTHDF2/miR-126 axis in AML and highlight the ther-apeutic potential of targeting this axis for AML treatment,but also suggest that m^(6)A plays a role in pre-miRNA processing that contributes to tumorigenesis.
