卵母细胞成熟过程受组蛋白H3K4me3(trimethylation of lysine 4 on histone 3)和H3K27me3(trimethylation of lysine 27 on histone 3)及其相关的甲基化和去甲基化酶的调控,因此考虑对鸡的卵泡发育也存在一定的影响。选取“苏禽3号”配...卵母细胞成熟过程受组蛋白H3K4me3(trimethylation of lysine 4 on histone 3)和H3K27me3(trimethylation of lysine 27 on histone 3)及其相关的甲基化和去甲基化酶的调控,因此考虑对鸡的卵泡发育也存在一定的影响。选取“苏禽3号”配套系第一母本为研究对象,采用Western blot法探究组蛋白H3K4me3和H3K27me3在鸡卵泡不同发育阶段颗粒层中蛋白的表达模式。结果表明:在苏禽3号卵泡颗粒层中,组蛋白H3K4me3在卵泡发育不同阶段表达模式呈降低→升高→降低→升高的波浪形趋势,波浪变化较为平缓,在F5、F2和F13个表达高点的表达量与SWF(small white follicle)、LWF(large white follicle)、SYF(small yellow follicle)和F34个表达低点的表达差异显著(P<0.05)。组蛋白H3K27me3在不同发育阶段表达模式亦呈波浪形表达趋势,波浪变化起伏较明显,在SWF、SYF和F33个表达高点的表达量与F5、F4、F1和F24个表达低点的表达差异显著(P<0.05)。相关性分析显示,组蛋白H3K4me3与H3K27me3在不同发育阶段卵泡颗粒细胞中的表达呈较强的负线性相关(R=-0.808,P=0.000)。结果提示:组蛋白H3K4me3和H3K27me3在不同发育阶段卵泡颗粒层中的表达具有组织差异性,呈负相关的动态修饰性,可能共同协调卵泡生长过程中各基因的表达与功能,研究结果为鸡繁殖性状调控机理提供了理论依据。展开更多
目的:探讨检测肝癌组织中组蛋白第三亚基四号赖氨酸的三甲基化(H3K4me3)蛋白的表达与肿瘤病理特点和肝癌患者生存预后的相关性。方法:免疫组化和Western-blot检测H3K4me3和组蛋白甲基转移酶(SET and MYND domain-containing protein 3,S...目的:探讨检测肝癌组织中组蛋白第三亚基四号赖氨酸的三甲基化(H3K4me3)蛋白的表达与肿瘤病理特点和肝癌患者生存预后的相关性。方法:免疫组化和Western-blot检测H3K4me3和组蛋白甲基转移酶(SET and MYND domain-containing protein 3,SMYD3)在肝癌组织(n=168)和细胞株中的表达。此外,实验结果还在另外一个肝癌组织芯片(n=147)中进行验证。H3K4me3表达的最佳分界点(optimal cut-point)由X-tile程序确定,患者的预后由Kaplan-meier生存曲线描述。结果:H3K4me3高表达于肝癌细胞系和肝癌组织,其高表达与肝癌尤其是早期TNM1/2期患者的较差总体生存显著相关。单因素和多因素分析均提示H3K4me3表达水平是患者预后的独立危险因素。此外,H3K4me3和SMYD3在两组肝癌组织中均存在正相关表达。结论:H3K4me3表达水平能成为肝癌患者术后生存的预测因子,其高表达可能与SMYD3有关。展开更多
Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such inf...Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.展开更多
Gastrokine 1 (GKN1) is a highly secreted gastric mucosal protein in normal individuals but strongly down-regulated or totally absent in gastric cancer subjects. An epigenetic mechanism might be responsible for GKN1 ge...Gastrokine 1 (GKN1) is a highly secreted gastric mucosal protein in normal individuals but strongly down-regulated or totally absent in gastric cancer subjects. An epigenetic mechanism might be responsible for GKN1 gene silencing probably through the activity of a transcription factor in association with the enzymes SUV39H1 and HDACs on the GKN1 promoter. In fact, compared to non-tumor tissues, a high increase of H3K9me3 level was observed in the corresponding tumor ones. Because H3K4me3 seems to be a possible epigenetic mark for active euchromatin, we try to verify the H3K4me3 level on the GKN1 promoter in gastric cancer tumor specimens. In addition, we also attempt to highlight if CBX7 could be the possible regulatory transcription factor correlated to GKN1 gene promoter. Therefore, we evaluated if the CBX7 expression levels could be associated with GKN1 down-regulation in gastric cancer. To this purpose, 2 pairs of non-tumor and tumor surgical specimens from patients with gastric cancer were analyzed for H3K4me3 by chromatin immunoprecipitation (ChiP) assays, and 9 pairs were instead analyzed by Western blotting for GKN1, and CBX7 expression levels, respectively. The results suggested that the observed increase of H3K4me3 in tumor samples was not in agreement with its proposed function whereas the expression of CBX7 was not associated with the down-regulation of GKN1. In particular, the expression levels of CBX7 in tumor samples might suggest a survival role in gastric cancer.展开更多
Histone H3 lysine 4 trimethylation(H3K4me3)is a canonical chromatin modification associated with active gene transcription,playing a pivotal role in regulating various cellular functions.Components of the H3K4me3 meth...Histone H3 lysine 4 trimethylation(H3K4me3)is a canonical chromatin modification associated with active gene transcription,playing a pivotal role in regulating various cellular functions.Components of the H3K4me3 methyltransferase complex,known as the proteins associated with SET1(COMPASS),have been implicated in exerting cancer-protective or cancer-inhibitory effects through inducive H3K4me3 modification.However,the role of the indispensable non-catalytic component of COMPASS CXXC-type zinc finger protein 1(CFP1)in malignant progression remains unclear.展开更多
The breadth of the enrichment site for post-translational trimethylation of histone H3 at lysine 4(H3K4me3)on chromatin has attracted great attention recently.H3K4me3,an extensively-studied histone modification,is rep...The breadth of the enrichment site for post-translational trimethylation of histone H3 at lysine 4(H3K4me3)on chromatin has attracted great attention recently.H3K4me3,an extensively-studied histone modification,is reported to promote gene transcription by directing preinitiation complex assembly through interaction with effector proteins,e.g.,the transcription factor IID(TFIID)complex[1].Scientists展开更多
Objective To observe the effect of arsenic exposure to drinking water on the level of histone 3 lysine 4 trimethylation(H3K4me3)and histone 3 lysine 79 trimethylation(H3K79me3)in peripheral blood leukocytes of human,a...Objective To observe the effect of arsenic exposure to drinking water on the level of histone 3 lysine 4 trimethylation(H3K4me3)and histone 3 lysine 79 trimethylation(H3K79me3)in peripheral blood leukocytes of human,and to analyze the relationship between arsenic exposure and H3K4me3,H3K79me3 modification levels.Methods A cluster sampling survey was carried out in typical endemic arsenicosis areas of Shanxi and Jilin provinces.Two hundred eighty-one local residents with a drinking water age of≥10 years were selected as the survey subjects.According to the arsenic content of drinking water,the tested population was divided into control group(water arsenic content≤0.01 mg/L,60 cases),low water arsenic exposure group(>0.01-0.05 mg/L,61 cases),medium water arsenic exposure group(>0.05-0.10 mg/L,50 cases),and 110 cases of high water arsenic exposure group(>0.10 mg/L).Drinking water samples,immediate urine samples and peripheral blood samples were collected from the subjects.Arsenic content in drinking water and urinary arsenic content were determined via the atomic fluorescence method;histone H3K4me3 and H3K79me3 in peripheral blood leukocytes were determined by dot blot hybridization(Dot Blotting).Results There were no statistically significant differences in age(61.50,60.00,59.50,59.50 years old),different gender(male:20,27,17,40 cases,female:40,34,33,70 cases),body mass index(BMI),smoking and drinking status between the control group,low,medium and high water arsenic exposure groups.Water arsenic content in the control group,low,medium and high water arsenic exposure groups(median:0.005,0.024,0.076,0.150 mg/L),urinary arsenic content(0.011,0.018,0.061,0.134 mg/L),and water arsenic cumulative exposure levels(0.342,1.641,5.273,7.716 mg)were compared between the groups,the differences were statistically significant(H=256.041,88.615,218.610,P<0.01).In the control group,low,medium and high water arsenic exposure groups,the modification levels of H3K4me3(0.100,0.059,0.083,0.083)and H3K79me3(0.049,0.036,0.055,0.052)in peripheral blood leukocytes were not significantly different(H=1.488,2.097,P>0.05).The levels of H3K4me3 and H3K79me3 in peripheral blood leukocytes were positively correlated with water arsenic content,urinary arsenic content,water arsenic cumulative exposure levels(r=0.245,0.221;0.299,0.318;0.149,0.149;P<0.01 or<0.05);there was a positive correlation between H3K4me3 and H3K79me3 modification levels(r=0.811,P<0.01).Conclusion There is a positive correlation between arsenic exposure through drinking water and the levels of H3K4me3 and H3K79me3 in the peripheral blood leukocytes of the population,but it is necessary to expand the sample size for further study.展开更多
Due to the difficulty in accurately identifying structural variants(SVs) across genomes,their impact on cisregulato ry diverge n ce of closely related species,especially fish,remains to be explored.Recently identified...Due to the difficulty in accurately identifying structural variants(SVs) across genomes,their impact on cisregulato ry diverge n ce of closely related species,especially fish,remains to be explored.Recently identified broad H3K4me3 domains are essential for the regulation of genes involved in several biological processes.However,the role of broad H3K4me3 domains in phenotypic divergence remains poorly understood.Siniperca chuatsi and S.scherzeri are closely related but divergent in several phenotypic traits,making them an ideal model to study cis-regulatory evolution in sister species.Here,we generated chromosome-level genomes of S.chuatsi and S.scherzeri,with assembled genome sizes of 716.35 and740.54 Mb,respectively.The evolutionary histories of S.chuatsi and S.scherzeri were studied by inferring dynamic changes in ancestral population sizes.To explore the genetic basis of adaptation in S.chuatsi and S.scherzeri,we performed gene family expansion and contraction analysis and identified positively selected genes(PSGs).To investigate the role of SVs in cis-regulatory divergence of closely related fish species,we identified high-quality SVs as well as divergent H3K27ac and H3K4me3 domains in the genomes of S.chuatsi and S.scherzeri.Integrated analysis revealed that cis-regulatory divergence caused by SVs played an essential role in phenotypic divergence between S.chuatsi and S.scherzeri.Additionally,divergent broad H3K4me3 domains were mostly associated with cancer-related genes in S.chuatsi and S.scherzeri and contributed to their phenotypic divergence.展开更多
文摘卵母细胞成熟过程受组蛋白H3K4me3(trimethylation of lysine 4 on histone 3)和H3K27me3(trimethylation of lysine 27 on histone 3)及其相关的甲基化和去甲基化酶的调控,因此考虑对鸡的卵泡发育也存在一定的影响。选取“苏禽3号”配套系第一母本为研究对象,采用Western blot法探究组蛋白H3K4me3和H3K27me3在鸡卵泡不同发育阶段颗粒层中蛋白的表达模式。结果表明:在苏禽3号卵泡颗粒层中,组蛋白H3K4me3在卵泡发育不同阶段表达模式呈降低→升高→降低→升高的波浪形趋势,波浪变化较为平缓,在F5、F2和F13个表达高点的表达量与SWF(small white follicle)、LWF(large white follicle)、SYF(small yellow follicle)和F34个表达低点的表达差异显著(P<0.05)。组蛋白H3K27me3在不同发育阶段表达模式亦呈波浪形表达趋势,波浪变化起伏较明显,在SWF、SYF和F33个表达高点的表达量与F5、F4、F1和F24个表达低点的表达差异显著(P<0.05)。相关性分析显示,组蛋白H3K4me3与H3K27me3在不同发育阶段卵泡颗粒细胞中的表达呈较强的负线性相关(R=-0.808,P=0.000)。结果提示:组蛋白H3K4me3和H3K27me3在不同发育阶段卵泡颗粒层中的表达具有组织差异性,呈负相关的动态修饰性,可能共同协调卵泡生长过程中各基因的表达与功能,研究结果为鸡繁殖性状调控机理提供了理论依据。
文摘目的:探讨检测肝癌组织中组蛋白第三亚基四号赖氨酸的三甲基化(H3K4me3)蛋白的表达与肿瘤病理特点和肝癌患者生存预后的相关性。方法:免疫组化和Western-blot检测H3K4me3和组蛋白甲基转移酶(SET and MYND domain-containing protein 3,SMYD3)在肝癌组织(n=168)和细胞株中的表达。此外,实验结果还在另外一个肝癌组织芯片(n=147)中进行验证。H3K4me3表达的最佳分界点(optimal cut-point)由X-tile程序确定,患者的预后由Kaplan-meier生存曲线描述。结果:H3K4me3高表达于肝癌细胞系和肝癌组织,其高表达与肝癌尤其是早期TNM1/2期患者的较差总体生存显著相关。单因素和多因素分析均提示H3K4me3表达水平是患者预后的独立危险因素。此外,H3K4me3和SMYD3在两组肝癌组织中均存在正相关表达。结论:H3K4me3表达水平能成为肝癌患者术后生存的预测因子,其高表达可能与SMYD3有关。
基金We thank the Experimental Poultry Farm of the Poultry Institute,Chinese Academy of Agricultural Sciences,for providing experimental materialsThis work was supported by the Key Research and Development Program of China(2017YFE0108000)+1 种基金the National Natural Science Foundation of China(31872341,31572390)the High-Level Talent Support Program of Yangzhou University,China.
文摘Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.
文摘Gastrokine 1 (GKN1) is a highly secreted gastric mucosal protein in normal individuals but strongly down-regulated or totally absent in gastric cancer subjects. An epigenetic mechanism might be responsible for GKN1 gene silencing probably through the activity of a transcription factor in association with the enzymes SUV39H1 and HDACs on the GKN1 promoter. In fact, compared to non-tumor tissues, a high increase of H3K9me3 level was observed in the corresponding tumor ones. Because H3K4me3 seems to be a possible epigenetic mark for active euchromatin, we try to verify the H3K4me3 level on the GKN1 promoter in gastric cancer tumor specimens. In addition, we also attempt to highlight if CBX7 could be the possible regulatory transcription factor correlated to GKN1 gene promoter. Therefore, we evaluated if the CBX7 expression levels could be associated with GKN1 down-regulation in gastric cancer. To this purpose, 2 pairs of non-tumor and tumor surgical specimens from patients with gastric cancer were analyzed for H3K4me3 by chromatin immunoprecipitation (ChiP) assays, and 9 pairs were instead analyzed by Western blotting for GKN1, and CBX7 expression levels, respectively. The results suggested that the observed increase of H3K4me3 in tumor samples was not in agreement with its proposed function whereas the expression of CBX7 was not associated with the down-regulation of GKN1. In particular, the expression levels of CBX7 in tumor samples might suggest a survival role in gastric cancer.
基金This work was supported by the National Key R&D Program of China(2021YFF1201303)the National Natural Science Foundation of China(81972196)+1 种基金the CAMS Innovation Fund for Medical Sciences(CIFMS)(2021-1-12M-012)the R&D Program of Beijing Municipal Education Commission(KJZD20191002302).
文摘Histone H3 lysine 4 trimethylation(H3K4me3)is a canonical chromatin modification associated with active gene transcription,playing a pivotal role in regulating various cellular functions.Components of the H3K4me3 methyltransferase complex,known as the proteins associated with SET1(COMPASS),have been implicated in exerting cancer-protective or cancer-inhibitory effects through inducive H3K4me3 modification.However,the role of the indispensable non-catalytic component of COMPASS CXXC-type zinc finger protein 1(CFP1)in malignant progression remains unclear.
基金supported by faculty start up funding provided by The Methodist Hospital Research Institute,Texas,United States
文摘The breadth of the enrichment site for post-translational trimethylation of histone H3 at lysine 4(H3K4me3)on chromatin has attracted great attention recently.H3K4me3,an extensively-studied histone modification,is reported to promote gene transcription by directing preinitiation complex assembly through interaction with effector proteins,e.g.,the transcription factor IID(TFIID)complex[1].Scientists
文摘Objective To observe the effect of arsenic exposure to drinking water on the level of histone 3 lysine 4 trimethylation(H3K4me3)and histone 3 lysine 79 trimethylation(H3K79me3)in peripheral blood leukocytes of human,and to analyze the relationship between arsenic exposure and H3K4me3,H3K79me3 modification levels.Methods A cluster sampling survey was carried out in typical endemic arsenicosis areas of Shanxi and Jilin provinces.Two hundred eighty-one local residents with a drinking water age of≥10 years were selected as the survey subjects.According to the arsenic content of drinking water,the tested population was divided into control group(water arsenic content≤0.01 mg/L,60 cases),low water arsenic exposure group(>0.01-0.05 mg/L,61 cases),medium water arsenic exposure group(>0.05-0.10 mg/L,50 cases),and 110 cases of high water arsenic exposure group(>0.10 mg/L).Drinking water samples,immediate urine samples and peripheral blood samples were collected from the subjects.Arsenic content in drinking water and urinary arsenic content were determined via the atomic fluorescence method;histone H3K4me3 and H3K79me3 in peripheral blood leukocytes were determined by dot blot hybridization(Dot Blotting).Results There were no statistically significant differences in age(61.50,60.00,59.50,59.50 years old),different gender(male:20,27,17,40 cases,female:40,34,33,70 cases),body mass index(BMI),smoking and drinking status between the control group,low,medium and high water arsenic exposure groups.Water arsenic content in the control group,low,medium and high water arsenic exposure groups(median:0.005,0.024,0.076,0.150 mg/L),urinary arsenic content(0.011,0.018,0.061,0.134 mg/L),and water arsenic cumulative exposure levels(0.342,1.641,5.273,7.716 mg)were compared between the groups,the differences were statistically significant(H=256.041,88.615,218.610,P<0.01).In the control group,low,medium and high water arsenic exposure groups,the modification levels of H3K4me3(0.100,0.059,0.083,0.083)and H3K79me3(0.049,0.036,0.055,0.052)in peripheral blood leukocytes were not significantly different(H=1.488,2.097,P>0.05).The levels of H3K4me3 and H3K79me3 in peripheral blood leukocytes were positively correlated with water arsenic content,urinary arsenic content,water arsenic cumulative exposure levels(r=0.245,0.221;0.299,0.318;0.149,0.149;P<0.01 or<0.05);there was a positive correlation between H3K4me3 and H3K79me3 modification levels(r=0.811,P<0.01).Conclusion There is a positive correlation between arsenic exposure through drinking water and the levels of H3K4me3 and H3K79me3 in the peripheral blood leukocytes of the population,but it is necessary to expand the sample size for further study.
基金supported by the National Natural Science Foundation of China (31900309)Guangdong Basic and Applied Basic Research Foundation (2019A1515011644)+2 种基金Key-Area Research and Development Program of Guangdong Province (2021B0202020001)Seed Industry Development Project of Agricultural and Rural Department of Guangdong Province (2022)Innovation Group Project of Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai)(311021006)。
文摘Due to the difficulty in accurately identifying structural variants(SVs) across genomes,their impact on cisregulato ry diverge n ce of closely related species,especially fish,remains to be explored.Recently identified broad H3K4me3 domains are essential for the regulation of genes involved in several biological processes.However,the role of broad H3K4me3 domains in phenotypic divergence remains poorly understood.Siniperca chuatsi and S.scherzeri are closely related but divergent in several phenotypic traits,making them an ideal model to study cis-regulatory evolution in sister species.Here,we generated chromosome-level genomes of S.chuatsi and S.scherzeri,with assembled genome sizes of 716.35 and740.54 Mb,respectively.The evolutionary histories of S.chuatsi and S.scherzeri were studied by inferring dynamic changes in ancestral population sizes.To explore the genetic basis of adaptation in S.chuatsi and S.scherzeri,we performed gene family expansion and contraction analysis and identified positively selected genes(PSGs).To investigate the role of SVs in cis-regulatory divergence of closely related fish species,we identified high-quality SVs as well as divergent H3K27ac and H3K4me3 domains in the genomes of S.chuatsi and S.scherzeri.Integrated analysis revealed that cis-regulatory divergence caused by SVs played an essential role in phenotypic divergence between S.chuatsi and S.scherzeri.Additionally,divergent broad H3K4me3 domains were mostly associated with cancer-related genes in S.chuatsi and S.scherzeri and contributed to their phenotypic divergence.
