DNA methylation is a critical epigenetic mechanism that influences gene transcription, genomic stability, X-chromosome inactivation and other factors, and appropriate DNA methylation is crucial in development. DNA met...DNA methylation is a critical epigenetic mechanism that influences gene transcription, genomic stability, X-chromosome inactivation and other factors, and appropriate DNA methylation is crucial in development. DNA methyltransferase 1 （DNMT1） plays an important role in maintaining the established methylation pattern during DNA replication. Although the effect of DNA methylation on embryonic development has been well known in vertebrates, little research has been carried out in invertebrates, especially in marine bivalves. In this study, the DNMT1 gene （MyDNMT1） was firstly identified from Mizuhopecten yessoensis. The full-length cDNA of MyDNMT1 was 5 039 bp, consisted of a 5＇ untranslated region （5＇-UTR） of 79 bp, a 3＇ untranslated region （3＇-UTR） of 199 bp, and a 4 761 bp open reading frame （ORF） encoding a peptide of 1 586 amino acids without a putative signal peptide. The relative mRNA expression level of MyDNMT1 was measured during the embryonic development of M. ydssoensis using real-time PCR, which revealed that the level at stage zygote and trochophore were significantly higher than that at other stages. We further examined the global DNA methylation during development by colorimetric method. The results showed that the methylation level was increased and reached the peak at blastula stage, then dramatically decreased, and fluctuated at early D-shaped larva stage. This study provided greater insight into the DNA methylation of embryonic development, which obtained a better understanding of the relationship between the DNA methylation and the embryonic development in bivalve mollusks.展开更多
目的研究当归芍药散对载脂蛋白E基因敲除(ApoE^(-/-))小鼠主动脉粥样硬化(atherosclerosis,As)斑块的干预作用及DNA甲基转移酶1(DNMT1)和PPAR-γ表达的影响。方法将30只8周龄雄性ApoE^(-/-)小鼠西方类型膳食喂养9周,随机分为模型组、立...目的研究当归芍药散对载脂蛋白E基因敲除(ApoE^(-/-))小鼠主动脉粥样硬化(atherosclerosis,As)斑块的干预作用及DNA甲基转移酶1(DNMT1)和PPAR-γ表达的影响。方法将30只8周龄雄性ApoE^(-/-)小鼠西方类型膳食喂养9周,随机分为模型组、立普妥组(阳性对照组)和当归芍药散组(n=10),药物干预9周后,检测血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、血清DNA甲基化水平及DNMTs水平。行苏木素-伊红染色(hematoxylin and eosin staining,HE),采用IPP图像分析软件测量小鼠主动脉As斑块面积。免疫组化法检测各组小鼠主动脉As斑块内DNA甲基化转移酶1(DNMT1)和过氧化物酶体增殖物活化受体γ(PPARγ)的表达。结果与模型组相比,当归芍药散组和立普妥组小鼠血清TC、TG水平明显降低(P<0.05),当归芍药散组小鼠血清HDL-C水平明显升高(P<0.05)。与模型组相比,当归芍药散组小鼠主动脉As斑块面积显著降低(P<0.01),当归芍药散组小鼠主动脉斑块内DNMT1表达显著降低(P<0.05),PPARγ表达显著升高(P<0.05)。结论当归芍药散可改善As小鼠的血脂水平,减少As斑块面积,具有明确的抗As作用,其机制可能与抑制As小鼠血清甲基化水平和DNMTs水平,促进As小鼠主动脉斑块内PPARγ表达和抑制斑块内DNMT1表达有关。展开更多
Objective To investigate the effects of DNA methyl transferase 1(DNMT1)gene regulation by miR 34a in osteosarcoma(OS)MG63 cells and to determine the possible mechanisms.Methods A miR-34a mimic,mimic negative control(N...Objective To investigate the effects of DNA methyl transferase 1(DNMT1)gene regulation by miR 34a in osteosarcoma(OS)MG63 cells and to determine the possible mechanisms.Methods A miR-34a mimic,mimic negative control(NC),miR-34a inhibitor,or inhibitor-NC were transfected into MG63 cells using Lipofectamine 3000.Cell viability was measured using the MTT assay.The mRNA levels of DNMT1 in MG63 cells were detected by qPCR.The proliferation of MG63 cells was detected by the CCK8 method.The protein expression of DNMT1 was measured by Western blotting.The effects of DNMT1 on the migration of MG63 cells were examined by a cell migration assay.The apoptosis of MG63 cells was detected by flow cytometry.Results The mRNA levels of DNMT1 were up-regulated in MG63 cells treated with the miR-34a inhibitor compared to other groups.The growth of MG63 cells was significantly increased in these cells.The protein expression of DNMT1 and the cell proliferation and migration decreased in the miR-34a mimic-treated group compared to the cells treated with the miR-34a inhibitor or the negative control(both P<0.05).Conclusion By targeting the DNMT1 gene,miR-34a reduces the expression level of DNMT1 in MG63 cells,thereby affecting the viability,migration,proliferation,and apoptosis of MG63 cells.Interventions targeting the miR-34a/DNMT1 axis may represent a novel targeted therapy for OS.展开更多
Associations between 3D chromatin architectures and epigenetic modifications have been characterized in animals.However,any impact of DNA methylation on chromatin architecture in plants is understudied,which is confin...Associations between 3D chromatin architectures and epigenetic modifications have been characterized in animals.However,any impact of DNA methylation on chromatin architecture in plants is understudied,which is confined to Arabidopsis thaliana.Because plant species differ in genome size,composition,and overall chromatin packing,it is unclear to what extent findings from A.thaliana hold in other species.Moreover,the incomplete chromatin architectural profiles and the low-resolution high-throughput chromosome conformation capture(Hi-C)data from A.thaliana have hampered characterizing its subtle chromatin structures and their associations with DNA methylation.We constructed a high-resolution Hi-C interaction map for the null OsMET1-2(the major CG methyltransferase in rice)mutant(osmet1-2)and isogenic wild-type rice(WT).Chromatin structural changes occurred in osmetl-2,including intra-/inter-chromosomal interactions,compartment transition,and topologically associated domains(TAD)variations.Our findings provide novel insights into the potential function of DNA methylation in TAD formation in rice and confirmed DNA methylation plays similar essential roles in chromatin packing in A,thaliana and rice.展开更多
基金Supported by Earmarked Fund for Modern Agro-industry Technology Research System of China(CARS-49)Natural Science Foundation of Liaoning Province(201602408)~~
文摘DNA methylation is a critical epigenetic mechanism that influences gene transcription, genomic stability, X-chromosome inactivation and other factors, and appropriate DNA methylation is crucial in development. DNA methyltransferase 1 （DNMT1） plays an important role in maintaining the established methylation pattern during DNA replication. Although the effect of DNA methylation on embryonic development has been well known in vertebrates, little research has been carried out in invertebrates, especially in marine bivalves. In this study, the DNMT1 gene （MyDNMT1） was firstly identified from Mizuhopecten yessoensis. The full-length cDNA of MyDNMT1 was 5 039 bp, consisted of a 5＇ untranslated region （5＇-UTR） of 79 bp, a 3＇ untranslated region （3＇-UTR） of 199 bp, and a 4 761 bp open reading frame （ORF） encoding a peptide of 1 586 amino acids without a putative signal peptide. The relative mRNA expression level of MyDNMT1 was measured during the embryonic development of M. ydssoensis using real-time PCR, which revealed that the level at stage zygote and trochophore were significantly higher than that at other stages. We further examined the global DNA methylation during development by colorimetric method. The results showed that the methylation level was increased and reached the peak at blastula stage, then dramatically decreased, and fluctuated at early D-shaped larva stage. This study provided greater insight into the DNA methylation of embryonic development, which obtained a better understanding of the relationship between the DNA methylation and the embryonic development in bivalve mollusks.
文摘目的研究当归芍药散对载脂蛋白E基因敲除(ApoE^(-/-))小鼠主动脉粥样硬化(atherosclerosis,As)斑块的干预作用及DNA甲基转移酶1(DNMT1)和PPAR-γ表达的影响。方法将30只8周龄雄性ApoE^(-/-)小鼠西方类型膳食喂养9周,随机分为模型组、立普妥组(阳性对照组)和当归芍药散组(n=10),药物干预9周后,检测血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、血清DNA甲基化水平及DNMTs水平。行苏木素-伊红染色(hematoxylin and eosin staining,HE),采用IPP图像分析软件测量小鼠主动脉As斑块面积。免疫组化法检测各组小鼠主动脉As斑块内DNA甲基化转移酶1(DNMT1)和过氧化物酶体增殖物活化受体γ(PPARγ)的表达。结果与模型组相比,当归芍药散组和立普妥组小鼠血清TC、TG水平明显降低(P<0.05),当归芍药散组小鼠血清HDL-C水平明显升高(P<0.05)。与模型组相比,当归芍药散组小鼠主动脉As斑块面积显著降低(P<0.01),当归芍药散组小鼠主动脉斑块内DNMT1表达显著降低(P<0.05),PPARγ表达显著升高(P<0.05)。结论当归芍药散可改善As小鼠的血脂水平,减少As斑块面积,具有明确的抗As作用,其机制可能与抑制As小鼠血清甲基化水平和DNMTs水平,促进As小鼠主动脉斑块内PPARγ表达和抑制斑块内DNMT1表达有关。
基金supported by grants from the Natural Science Foundation of Fujian Province(No.2019J01011 to CY Jia)the Open Project of Key Laboratory of Union Hospital Affiliated to Fujian Medical University(No.XHZDSYS202004 to P Wang).
文摘Objective To investigate the effects of DNA methyl transferase 1(DNMT1)gene regulation by miR 34a in osteosarcoma(OS)MG63 cells and to determine the possible mechanisms.Methods A miR-34a mimic,mimic negative control(NC),miR-34a inhibitor,or inhibitor-NC were transfected into MG63 cells using Lipofectamine 3000.Cell viability was measured using the MTT assay.The mRNA levels of DNMT1 in MG63 cells were detected by qPCR.The proliferation of MG63 cells was detected by the CCK8 method.The protein expression of DNMT1 was measured by Western blotting.The effects of DNMT1 on the migration of MG63 cells were examined by a cell migration assay.The apoptosis of MG63 cells was detected by flow cytometry.Results The mRNA levels of DNMT1 were up-regulated in MG63 cells treated with the miR-34a inhibitor compared to other groups.The growth of MG63 cells was significantly increased in these cells.The protein expression of DNMT1 and the cell proliferation and migration decreased in the miR-34a mimic-treated group compared to the cells treated with the miR-34a inhibitor or the negative control(both P<0.05).Conclusion By targeting the DNMT1 gene,miR-34a reduces the expression level of DNMT1 in MG63 cells,thereby affecting the viability,migration,proliferation,and apoptosis of MG63 cells.Interventions targeting the miR-34a/DNMT1 axis may represent a novel targeted therapy for OS.
基金the National Natural Science Foundation of China(Grant No.31670220)the Program of Changbai Mountain Scholar。
文摘Associations between 3D chromatin architectures and epigenetic modifications have been characterized in animals.However,any impact of DNA methylation on chromatin architecture in plants is understudied,which is confined to Arabidopsis thaliana.Because plant species differ in genome size,composition,and overall chromatin packing,it is unclear to what extent findings from A.thaliana hold in other species.Moreover,the incomplete chromatin architectural profiles and the low-resolution high-throughput chromosome conformation capture(Hi-C)data from A.thaliana have hampered characterizing its subtle chromatin structures and their associations with DNA methylation.We constructed a high-resolution Hi-C interaction map for the null OsMET1-2(the major CG methyltransferase in rice)mutant(osmet1-2)and isogenic wild-type rice(WT).Chromatin structural changes occurred in osmetl-2,including intra-/inter-chromosomal interactions,compartment transition,and topologically associated domains(TAD)variations.Our findings provide novel insights into the potential function of DNA methylation in TAD formation in rice and confirmed DNA methylation plays similar essential roles in chromatin packing in A,thaliana and rice.
