Background:Epigenetic regulation of the gene expression results from interaction between the external environment and transcription of the genetic information encoded in DNA.Methylated CpG regions within the gene prom...Background:Epigenetic regulation of the gene expression results from interaction between the external environment and transcription of the genetic information encoded in DNA.Methylated CpG regions within the gene promoters lead to silencing of the gene expression in most cases.Factors contributing to epigenetic regulation include intestinal microbiota,which in chicken can be potently modified by in ovo stimulation.The main aim of this study was to determine global and specific methylation patterns of the spleen under the influence of host-microbiome interaction.Results:Fertilized eggs of two genotypes:Ross 308 and Green-legged Partridgelike were in ovo stimulated on d 12 of incubation.The injected compounds were as follows:probiotic-Lactococcus lactis subsp.cremoris IBB477,prebiotic-galactooligosaccharides,and synbiotic-combination of both.Chickens were sacrificed on d 42 post-hatching.Spleen was collected,RNA and DNA were isolated and intended to gene expression,gene methylation and global methylation analysis.We have proved that negative regulation of gene expression after administration of bioactive substances in ovo might have epigenetic character.Epigenetic changes depend on the genotype and the substance administered in ovo.Conclusion:Epigenetic nature of microbial reprogramming in poultry and extension of issues related to hostmicrobiome interaction is a new direction of this research.展开更多
目的探讨脑膜瘤mut L同源基因1（MLH1）启动子甲基化水平与脑膜瘤侵袭性的相关性。方法从我院标本库随机选取64例脑膜瘤石蜡包埋组织标本,其中侵袭性脑膜瘤26例,非侵袭性脑膜瘤38例。应用逆转录-PCR检测MLH1 m RNA表达水平,应用特异高...目的探讨脑膜瘤mut L同源基因1（MLH1）启动子甲基化水平与脑膜瘤侵袭性的相关性。方法从我院标本库随机选取64例脑膜瘤石蜡包埋组织标本,其中侵袭性脑膜瘤26例,非侵袭性脑膜瘤38例。应用逆转录-PCR检测MLH1 m RNA表达水平,应用特异高分辨率熔解（MS-HRM）曲线检测不同侵袭性脑膜瘤MLH1启动子甲基化水平。结果侵袭性脑膜瘤MLH1m RNA含量比非侵袭性脑膜瘤显著降低（P〈0.05）。MS-HRM曲线分析显示,侵袭性脑膜瘤MLH1启动子区高度甲基化水平为0-1%所占比例（7.7%,2/26）显著低于非侵袭性脑膜瘤（55.3%,21/38;P〈0.05）。结论 MLH1启动子区甲基化可能下调MLH1的表达,进而调控脑膜瘤侵袭性行为。展开更多
目的:探讨乳腺癌MDA-MB-231细胞中,Y性别决定区基因7(SOX7)基因启动子甲基化水平对细胞的体外迁移和侵袭的影响。方法:脂质体转染pcDNA3.0-DNA甲基转移酶3a(DNMT3a)质粒至MDA-MB-231细胞中,并于24h、48h及72h后,采用蛋白质免疫印迹实验(...目的:探讨乳腺癌MDA-MB-231细胞中,Y性别决定区基因7(SOX7)基因启动子甲基化水平对细胞的体外迁移和侵袭的影响。方法:脂质体转染pcDNA3.0-DNA甲基转移酶3a(DNMT3a)质粒至MDA-MB-231细胞中,并于24h、48h及72h后,采用蛋白质免疫印迹实验(WB)检测细胞内DNMT3a蛋白表达水平;甲基化特异性定量PCR(Q-MSP)检测DNMT3a处理组、5-aza-C处理组及对照(Control)组MDA-MB-231细胞中的SOX7基因启动子DNA甲基化水平;实时荧光定量PCR(qRT-PCR)及WB实验检测各组MDA-MB-231细胞中的SOX7 m RNA和蛋白表达水平;细胞划痕实验及细胞侵袭实验检测各组MDA-MB-231细胞的迁移和侵袭能力。结果:pcDNA3.0-DNMT3a质粒转染MDA-MB-231细胞24h时,细胞内的DNMT3a蛋白表达水平最高。DNMT3a能够显著提高SOX7基因启动子DNA甲基化水平,而5-aza-C则抑制了SOX7基因启动子DNA甲基化水平(P<0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞中,SOX7的m RNA及蛋白表达水平均明显下降,而5-aza-C处理组SOX7的m RNA及蛋白表达水平均明显增加(P<0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞的迁移和侵袭能力均显著增强(P<0.05),而5-aza-C处理组的MDA-MB-231细胞的迁移和侵袭能力变化不大(P>0.05)。结论:在恶性肿瘤中,SOX7低表达表受其基因启动子高甲基化调节,且乳腺癌MDA-MB-231细胞中低表达的SOX7能够影响细胞的外迁移和侵袭能力。展开更多
Telomerase plays an essential role in many biological processes.DNA methylation regulates the expression of many genes,including telomerase.Here,we propose a deformable satellite nanocapsule fluorescein isothiocyanate...Telomerase plays an essential role in many biological processes.DNA methylation regulates the expression of many genes,including telomerase.Here,we propose a deformable satellite nanocapsule fluorescein isothiocyanate(FITC)-hollowbowl mesoporous organicsilica@gold nanoparticles-methyl-CpG-binding protein 2(MECP 2)-silver nanoclusters(FHBMO@AMA),for simultaneous quantitative detection of both cytoplasmic telomerase activity and the degree of DNA methylation.This strategy enabled spatial-based detection in cells.The total cytoplasmic telomerase activity was detected by fluorescence energy resonance transfer(FRET)between FHBMO and gold nanoparticles(Au NPs),while the DNA methylation in the nucleus was detected by enhanced fluorescence of silver nanoclusters(Ag NCs).Furthermore,FHBMO@AMA could intuitively distinguish between the differences in telomerase expression in cells during the DNA synthesis period at the mitotic phase(S/M)of the cell cycle.Interestingly,the ratio of the two detections(telomerase activity/DNA methylation)significantly correlated with the efficacy of anticancer drugs.At the same time,there was no apparent linear relationship between any single detection target and the efficacy of the anticancer drugs.Therefore,based on the relationship between telomerase activity and DNA methylation,our newly developed approach serves as new and feasible method for evaluating the efficacy of anticancer drugs,thereby,extending the technology toolbox for precision in medical and pharmaceutical analysis of drug potency.展开更多
基金supported by grant UMO-2017/25/N/NZ9/01822 funded by the National Science Centre in Cracow(Poland).
文摘Background:Epigenetic regulation of the gene expression results from interaction between the external environment and transcription of the genetic information encoded in DNA.Methylated CpG regions within the gene promoters lead to silencing of the gene expression in most cases.Factors contributing to epigenetic regulation include intestinal microbiota,which in chicken can be potently modified by in ovo stimulation.The main aim of this study was to determine global and specific methylation patterns of the spleen under the influence of host-microbiome interaction.Results:Fertilized eggs of two genotypes:Ross 308 and Green-legged Partridgelike were in ovo stimulated on d 12 of incubation.The injected compounds were as follows:probiotic-Lactococcus lactis subsp.cremoris IBB477,prebiotic-galactooligosaccharides,and synbiotic-combination of both.Chickens were sacrificed on d 42 post-hatching.Spleen was collected,RNA and DNA were isolated and intended to gene expression,gene methylation and global methylation analysis.We have proved that negative regulation of gene expression after administration of bioactive substances in ovo might have epigenetic character.Epigenetic changes depend on the genotype and the substance administered in ovo.Conclusion:Epigenetic nature of microbial reprogramming in poultry and extension of issues related to hostmicrobiome interaction is a new direction of this research.
文摘目的:探讨乳腺癌MDA-MB-231细胞中,Y性别决定区基因7(SOX7)基因启动子甲基化水平对细胞的体外迁移和侵袭的影响。方法:脂质体转染pcDNA3.0-DNA甲基转移酶3a(DNMT3a)质粒至MDA-MB-231细胞中,并于24h、48h及72h后,采用蛋白质免疫印迹实验(WB)检测细胞内DNMT3a蛋白表达水平;甲基化特异性定量PCR(Q-MSP)检测DNMT3a处理组、5-aza-C处理组及对照(Control)组MDA-MB-231细胞中的SOX7基因启动子DNA甲基化水平;实时荧光定量PCR(qRT-PCR)及WB实验检测各组MDA-MB-231细胞中的SOX7 m RNA和蛋白表达水平;细胞划痕实验及细胞侵袭实验检测各组MDA-MB-231细胞的迁移和侵袭能力。结果:pcDNA3.0-DNMT3a质粒转染MDA-MB-231细胞24h时,细胞内的DNMT3a蛋白表达水平最高。DNMT3a能够显著提高SOX7基因启动子DNA甲基化水平,而5-aza-C则抑制了SOX7基因启动子DNA甲基化水平(P<0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞中,SOX7的m RNA及蛋白表达水平均明显下降,而5-aza-C处理组SOX7的m RNA及蛋白表达水平均明显增加(P<0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞的迁移和侵袭能力均显著增强(P<0.05),而5-aza-C处理组的MDA-MB-231细胞的迁移和侵袭能力变化不大(P>0.05)。结论:在恶性肿瘤中,SOX7低表达表受其基因启动子高甲基化调节,且乳腺癌MDA-MB-231细胞中低表达的SOX7能够影响细胞的外迁移和侵袭能力。
基金The authors gratefully acknowledge the financial support from the National Natural Science Foundation of China(nos.21834004 and 21904063)the Natural Science Foundation of Jiangsu Province(no.BK20190279).
文摘Telomerase plays an essential role in many biological processes.DNA methylation regulates the expression of many genes,including telomerase.Here,we propose a deformable satellite nanocapsule fluorescein isothiocyanate(FITC)-hollowbowl mesoporous organicsilica@gold nanoparticles-methyl-CpG-binding protein 2(MECP 2)-silver nanoclusters(FHBMO@AMA),for simultaneous quantitative detection of both cytoplasmic telomerase activity and the degree of DNA methylation.This strategy enabled spatial-based detection in cells.The total cytoplasmic telomerase activity was detected by fluorescence energy resonance transfer(FRET)between FHBMO and gold nanoparticles(Au NPs),while the DNA methylation in the nucleus was detected by enhanced fluorescence of silver nanoclusters(Ag NCs).Furthermore,FHBMO@AMA could intuitively distinguish between the differences in telomerase expression in cells during the DNA synthesis period at the mitotic phase(S/M)of the cell cycle.Interestingly,the ratio of the two detections(telomerase activity/DNA methylation)significantly correlated with the efficacy of anticancer drugs.At the same time,there was no apparent linear relationship between any single detection target and the efficacy of the anticancer drugs.Therefore,based on the relationship between telomerase activity and DNA methylation,our newly developed approach serves as new and feasible method for evaluating the efficacy of anticancer drugs,thereby,extending the technology toolbox for precision in medical and pharmaceutical analysis of drug potency.
