目的研究分析胎膜早破(PROM)并发绒毛膜羊膜炎患者胎盘组织中组蛋白H3第4位赖氨酸三甲基化(H3K4me3)、组蛋白H3第27位赖氨酸三甲基化(H3K27me3)及修饰酶的表达水平及其临床意义。方法回顾性收集2021年6月~2023年12月因PROM在西安高新医...目的研究分析胎膜早破(PROM)并发绒毛膜羊膜炎患者胎盘组织中组蛋白H3第4位赖氨酸三甲基化(H3K4me3)、组蛋白H3第27位赖氨酸三甲基化(H3K27me3)及修饰酶的表达水平及其临床意义。方法回顾性收集2021年6月~2023年12月因PROM在西安高新医院就诊的95例孕妇临床资料,根据是否并发组织学绒毛膜羊膜炎(HCA)分为感染组(n=41)和非感染组(n=54);另选取同期30例正常妊娠产妇作为对照组。取所有受试者胎盘组织,检测H3K4me3,H3K27me3的表达水平;实时荧光定量PCR(qRT-PCR)检测H3K4me3,H3K27me3特异性甲基化转移酶[混合谱系白血病蛋白1(MLL1),Zeste增强子同源物2(EZH2)]和去甲基化转移酶[赖氨酸特异性去甲基化酶5B(KDM5B),Jumonji结构域包含蛋白3(JMJD3)]水平。分析三组H3K4me3,H3K27me3及特异性修饰酶表达差异及与HCA组织学分期的相关性;绘制受试者工作特征(ROC)曲线,计算曲线下面积(AUC)值,评估H3K4me3,H3K27me3对并发HCA感染的预测价值。结果与对照组相比,感染组和非感染组H3K4me3(0.08%±0.02%,0.12%±0.03%vs 0.25%±0.06%)及MLL1 mRNA(1.32±0.16,1.44±0.23 vs 2.02±0.31),JMJD3 mRNA(0.78±0.13,0.91±0.15 vs 1.53±0.23)水平明显降低(t=10.939~19.062),H3K27me39(0.23%±0.05%,0.15%±0.03%vs 0.10%±0.02%)及EZH2 mRNA(1.96±0.26,1.85±0.25 vs 1.15±0.29),KDM5B mRNA(1.46±0.15,1.35±0.18 vs 0.94±0.12)水平明显升高(t=6.077~14.974),且感染组各水平低于/高于非感染组(t=2.017~10.688),差异具有统计学意义(均P<0.05)。PROM胎盘组织中H3K4me3与H3K27me3水平呈显著负相关(r=-0.604,P<0.05);H3K4me3,MLL1,JMJD3 mRNA水平分别与HCA组织学分期呈负相关(r=-0.646,-0.489,-0.503,均P<0.05);H3K27me3,EZH2 mRNA,KDM5B mRNA水平分别与HCA组织学分期呈正相关(r=0.632,0.515,0.520,均P<0.05)。当H3K4me3,H3K27me3截断值分别取0.10%,0.19%时,两者联合预测PROM并发HCA感染的AUC(95%CI)值为0.896(0.882~0.947),敏感度、特异度分别为92.14%和86.23%,明显高于两者单一指标诊断。结论PROM胎盘组织中H3K4me3,H3K27me3及修饰酶水平与HCA感染及其组织学分期密切相关,对并发HCA感染具有较高的临床预测价值,有望作为临床诊断的新型生物标志物。展开更多
目的描绘弥漫型胃癌组织中组蛋白H3第27位赖氨酸的三甲基化(H3K27me3)修饰的全基因组分布图谱,通过鉴定H3K27me3所调控的关键靶基因,初步探究H3K27me3修饰重编程可能调控弥漫型胃癌细胞发生发展的作用机制。方法样本来源于2021-2023年...目的描绘弥漫型胃癌组织中组蛋白H3第27位赖氨酸的三甲基化(H3K27me3)修饰的全基因组分布图谱,通过鉴定H3K27me3所调控的关键靶基因,初步探究H3K27me3修饰重编程可能调控弥漫型胃癌细胞发生发展的作用机制。方法样本来源于2021-2023年在陆军特色医学中心消化内科内镜中心及手术室胃肠外科组接受检查或治疗的患者。共收集到正常组患者14例,其中男性6例,女性8例,平均年龄46岁;胃癌组患者14例,其中男性8例,女性6例,平均年龄63岁。采用染色质靶向剪切及转座酶技术(cleavage under target and tagmentation,CUT&Tag)捕获基因组H3K27me3修饰区域,分析H3K27me3修饰重编程特征。整合转录组(RNA‐Seq)测序数据、高通量染色体构象捕获技术(high‐throughput chromosome conformation capture,Hi‐C)及已发表的公共单细胞数据,分析H3K27me3修饰重编程在弥漫型胃癌细胞中所调控靶基因。结果CUT&Tag和RNA测序数据质量符合下游分析标准,正常胃黏膜组织和弥漫型胃癌组织的组蛋白H3K27me3修饰均主要分布于远端基因间区和内含子区。相较于正常组织,胃癌组织的H3K27me3修饰存在显著的重编程特征,表现为H3K27me3总体信号强度明显降低。其中缺失的2912个H3K27me3信号峰可能导致822个肿瘤相关基因的表达上调,这些基因中上调最显著(信号值强度的差异倍数≥2,P<0.05)的56个基因主要富集于哺乳动物雷帕霉素靶蛋白复合体1(mammalian target of rapamycin complex 1,mTORC1)信号通路,其中甲硫氨酸转运体SLC7A5和胱氨酸转运体SLC7A11在胃癌组织中的表达最高。单细胞数据提示,弥漫型胃癌组织中SLC7A11的异常高表达主要存在于肿瘤上皮细胞。利用公共数据和免疫组织化学实验进一步验证SLC7A11在弥漫型胃癌中高表达,且与胃癌患者的不良预后相关。结论组蛋白H3K27me3修饰重编程是弥漫型胃癌的重要表观遗传学特征;组蛋白H3K27me3修饰缺失可能上调肿瘤细胞SLC7A11表达,进而促进肿瘤进展。展开更多
The leaf is a major organ for photosynthesis,and its shape plays an important role in plant development and yield determination in rice(Oryza sativa L.).In this study,an adaxial curled leaf mutant,termed curly leaf 1-...The leaf is a major organ for photosynthesis,and its shape plays an important role in plant development and yield determination in rice(Oryza sativa L.).In this study,an adaxial curled leaf mutant,termed curly leaf 1-1(cul1-1),was obtained by chemical mutagenesis.The leaf rolling index of the cul1-1 mutant was higher than that of the wild-type,which was caused by the abnormal development of bulliform cells(BCs).We cloned the CUL1 gene by map-based cloning.A nonsense mutation was present in the cul1-1 mutant,converting a tryptophan codon into a stop codon.The CUL1 gene encodes a chromodomain,helicase/ATPase and DNA-binding domain containing protein.Genes related to leaf rolling and BC development,such as ADL1,REL1 and ROC5,were activated by the cul1-1 mutation.The trimethylation of lysine 27 in histone 3(H3K27me3),but not H3K4me3,at the ADL1,REL1 and ROC5 loci,was reduced in the cul1-1 mutant.High-throughput mRNA sequencing indicated that the cul1-1 mutation caused genome-wide differential gene expression.The differentially expressed genes were classified into a few gene ontology terms and Kyoto encyclopedia of genes and genomes pathways.In the natural population,22 missense genomic variations in the CUL1 locus were identified,which composed of 7 haplotypes.A haplotype network was also built with haplotype II as the ancestor.The findings revealed that CUL1 is essential for normal leaf development and regulates this process by inhibiting the expression of genes involved in leaf rolling and BC development.展开更多
文摘目的研究分析胎膜早破(PROM)并发绒毛膜羊膜炎患者胎盘组织中组蛋白H3第4位赖氨酸三甲基化(H3K4me3)、组蛋白H3第27位赖氨酸三甲基化(H3K27me3)及修饰酶的表达水平及其临床意义。方法回顾性收集2021年6月~2023年12月因PROM在西安高新医院就诊的95例孕妇临床资料,根据是否并发组织学绒毛膜羊膜炎(HCA)分为感染组(n=41)和非感染组(n=54);另选取同期30例正常妊娠产妇作为对照组。取所有受试者胎盘组织,检测H3K4me3,H3K27me3的表达水平;实时荧光定量PCR(qRT-PCR)检测H3K4me3,H3K27me3特异性甲基化转移酶[混合谱系白血病蛋白1(MLL1),Zeste增强子同源物2(EZH2)]和去甲基化转移酶[赖氨酸特异性去甲基化酶5B(KDM5B),Jumonji结构域包含蛋白3(JMJD3)]水平。分析三组H3K4me3,H3K27me3及特异性修饰酶表达差异及与HCA组织学分期的相关性;绘制受试者工作特征(ROC)曲线,计算曲线下面积(AUC)值,评估H3K4me3,H3K27me3对并发HCA感染的预测价值。结果与对照组相比,感染组和非感染组H3K4me3(0.08%±0.02%,0.12%±0.03%vs 0.25%±0.06%)及MLL1 mRNA(1.32±0.16,1.44±0.23 vs 2.02±0.31),JMJD3 mRNA(0.78±0.13,0.91±0.15 vs 1.53±0.23)水平明显降低(t=10.939~19.062),H3K27me39(0.23%±0.05%,0.15%±0.03%vs 0.10%±0.02%)及EZH2 mRNA(1.96±0.26,1.85±0.25 vs 1.15±0.29),KDM5B mRNA(1.46±0.15,1.35±0.18 vs 0.94±0.12)水平明显升高(t=6.077~14.974),且感染组各水平低于/高于非感染组(t=2.017~10.688),差异具有统计学意义(均P<0.05)。PROM胎盘组织中H3K4me3与H3K27me3水平呈显著负相关(r=-0.604,P<0.05);H3K4me3,MLL1,JMJD3 mRNA水平分别与HCA组织学分期呈负相关(r=-0.646,-0.489,-0.503,均P<0.05);H3K27me3,EZH2 mRNA,KDM5B mRNA水平分别与HCA组织学分期呈正相关(r=0.632,0.515,0.520,均P<0.05)。当H3K4me3,H3K27me3截断值分别取0.10%,0.19%时,两者联合预测PROM并发HCA感染的AUC(95%CI)值为0.896(0.882~0.947),敏感度、特异度分别为92.14%和86.23%,明显高于两者单一指标诊断。结论PROM胎盘组织中H3K4me3,H3K27me3及修饰酶水平与HCA感染及其组织学分期密切相关,对并发HCA感染具有较高的临床预测价值,有望作为临床诊断的新型生物标志物。
文摘目的描绘弥漫型胃癌组织中组蛋白H3第27位赖氨酸的三甲基化(H3K27me3)修饰的全基因组分布图谱,通过鉴定H3K27me3所调控的关键靶基因,初步探究H3K27me3修饰重编程可能调控弥漫型胃癌细胞发生发展的作用机制。方法样本来源于2021-2023年在陆军特色医学中心消化内科内镜中心及手术室胃肠外科组接受检查或治疗的患者。共收集到正常组患者14例,其中男性6例,女性8例,平均年龄46岁;胃癌组患者14例,其中男性8例,女性6例,平均年龄63岁。采用染色质靶向剪切及转座酶技术(cleavage under target and tagmentation,CUT&Tag)捕获基因组H3K27me3修饰区域,分析H3K27me3修饰重编程特征。整合转录组(RNA‐Seq)测序数据、高通量染色体构象捕获技术(high‐throughput chromosome conformation capture,Hi‐C)及已发表的公共单细胞数据,分析H3K27me3修饰重编程在弥漫型胃癌细胞中所调控靶基因。结果CUT&Tag和RNA测序数据质量符合下游分析标准,正常胃黏膜组织和弥漫型胃癌组织的组蛋白H3K27me3修饰均主要分布于远端基因间区和内含子区。相较于正常组织,胃癌组织的H3K27me3修饰存在显著的重编程特征,表现为H3K27me3总体信号强度明显降低。其中缺失的2912个H3K27me3信号峰可能导致822个肿瘤相关基因的表达上调,这些基因中上调最显著(信号值强度的差异倍数≥2,P<0.05)的56个基因主要富集于哺乳动物雷帕霉素靶蛋白复合体1(mammalian target of rapamycin complex 1,mTORC1)信号通路,其中甲硫氨酸转运体SLC7A5和胱氨酸转运体SLC7A11在胃癌组织中的表达最高。单细胞数据提示,弥漫型胃癌组织中SLC7A11的异常高表达主要存在于肿瘤上皮细胞。利用公共数据和免疫组织化学实验进一步验证SLC7A11在弥漫型胃癌中高表达,且与胃癌患者的不良预后相关。结论组蛋白H3K27me3修饰重编程是弥漫型胃癌的重要表观遗传学特征;组蛋白H3K27me3修饰缺失可能上调肿瘤细胞SLC7A11表达,进而促进肿瘤进展。
基金supported by the National Natural Science Foundation of China(32070642 and 31371222 to Dr.Xiaoxue Wang)the National Key Research and Development Program from the Ministry of Science and Technology of China(2016YFD0100406 and 2017YFD0300107 to Dr.Xiaoxue Wang)the Science and Technology Department of Liaoning province(2022JH6/100100039 to Dr.Xiaoxue Wang)。
文摘The leaf is a major organ for photosynthesis,and its shape plays an important role in plant development and yield determination in rice(Oryza sativa L.).In this study,an adaxial curled leaf mutant,termed curly leaf 1-1(cul1-1),was obtained by chemical mutagenesis.The leaf rolling index of the cul1-1 mutant was higher than that of the wild-type,which was caused by the abnormal development of bulliform cells(BCs).We cloned the CUL1 gene by map-based cloning.A nonsense mutation was present in the cul1-1 mutant,converting a tryptophan codon into a stop codon.The CUL1 gene encodes a chromodomain,helicase/ATPase and DNA-binding domain containing protein.Genes related to leaf rolling and BC development,such as ADL1,REL1 and ROC5,were activated by the cul1-1 mutation.The trimethylation of lysine 27 in histone 3(H3K27me3),but not H3K4me3,at the ADL1,REL1 and ROC5 loci,was reduced in the cul1-1 mutant.High-throughput mRNA sequencing indicated that the cul1-1 mutation caused genome-wide differential gene expression.The differentially expressed genes were classified into a few gene ontology terms and Kyoto encyclopedia of genes and genomes pathways.In the natural population,22 missense genomic variations in the CUL1 locus were identified,which composed of 7 haplotypes.A haplotype network was also built with haplotype II as the ancestor.The findings revealed that CUL1 is essential for normal leaf development and regulates this process by inhibiting the expression of genes involved in leaf rolling and BC development.
