Endometrial cancer （EC） is the most common and lethal gynaecological cancer type in Europe and in North America. Frequently EC arises more in the corpus proper and manifests as round, polypoid expansile masses, but ...Endometrial cancer （EC） is the most common and lethal gynaecological cancer type in Europe and in North America. Frequently EC arises more in the corpus proper and manifests as round, polypoid expansile masses, but it may also originate in the lower uterine segment or spread in endometrium with necrosis and hemorrhage. The analysis was performed using a custom panel containing all DNA sequences loci coding pre-miRNAs and genes related to biogenesis and regulation of sncRNAs in normal and tumor tissues extracted from 6 unrelated patients with endometrial carcinoma. The identified variations were correlated with mature miRNAs differentially expressed in the same normal and tumor endometrial tissues. The comparison analysis confirmed the high degree of cellular and genetic intratumoral heterogeneity with a temporal and spatial miRNA expression distribution in association with genomic variants identified. The classification of specific DNA mutations, onto the loci identified, should be suitable to characterize possible instability genome regions and help classification of tumors to ameliorate the clinical management of patients affected by endometrial carcinoma.展开更多
旨在筛选并验证调控牛增强子结合蛋白A(CCAAT/enhancer binding protein alpha,CEBPA)表达的miRNA及其对肌内前体脂肪细胞增殖和分化的影响。本研究综合运用多个在线软件筛选与CEBPA基因具有潜在调控关系的miRNA,运用qRT-PCR技术检测CE...旨在筛选并验证调控牛增强子结合蛋白A(CCAAT/enhancer binding protein alpha,CEBPA)表达的miRNA及其对肌内前体脂肪细胞增殖和分化的影响。本研究综合运用多个在线软件筛选与CEBPA基因具有潜在调控关系的miRNA,运用qRT-PCR技术检测CEBPA基因在关岭牛各类组织中的表达水平;构建CEBPA-3′UTR-WT和CEBPA-3′UTR-MUT双荧光素酶报告载体,通过双荧光素酶报告系统验证候选miRNA与CEBPA基因的结合关系;分离培养关岭牛肌内前体脂肪细胞,利用qRT-PCR、Western blot、油红O染色和CCK-8探究调控CEBPA的miRNA对牛肌内前体脂肪细胞分化与增殖的影响。生物信息学分析发现,bta-miR-23a、bta-miR-23b-3p、bta-miR-181a、bta-miR-181b、bta-miR-181c、bta-miR-181d与CEBPA基因具有潜在结合关系;qRT-PCR结果表明,CEBPA基因在关岭牛内脏组织、肌肉组织、脂肪、下丘脑以及部分生殖器官中均有表达,在2岁牛背最长肌的表达量极显著高于1日龄犊牛(P<0.01);双荧光素酶报告系统结果表明,bta-miR-23a、bta-miR-23b-3p、bta-miR-181a能直接结合牛CEBPA基因3′UTR;细胞试验结果表明,CEBPA基因在牛肌内前体脂肪细胞分化中的表达水平随时间逐渐升高,bta-miR-23a、bta-miR-23b-3p、bta-miR-181a在分化0 d表达水平极显著高于其他时期(P<0.01),整个分化时期bta-miR-23b-3p、bta-miR-181a与CEBPA基因表达趋势相反;过表达miR-23a、miR-23b-3p、miR-181a均能极显著降低CEBPA基因mRNA和蛋白表达量(P<0.01),并降低牛肌内前体脂肪细胞中脂滴含量;bta-miR-23a、bta-miR-23b-3p、bta-miR-181a能阶段性促进细胞增殖。结果表明,bta-miR-23a、bta-miR-23b-3p、bta-miR-181a能阶段性的促进牛肌内前体脂肪细胞增殖,并通过直接负调控CEBPA的表达来间接影响该细胞分化。展开更多
Curcumin exerts a neuroprotective effect on Alzheimer’s disease;however,it is not known whether microRNAs are involved in this protective effect.This study was conducted using swAPP695-HEK293 cells as an Alzheimer’s...Curcumin exerts a neuroprotective effect on Alzheimer’s disease;however,it is not known whether microRNAs are involved in this protective effect.This study was conducted using swAPP695-HEK293 cells as an Alzheimer’s disease cell model.swAPP695-HEK293 cells were treated with 0,0.5,1,2,5,and 10μM curcumin for 24 hours.The changes in miR-15b-5p,miR-19a-3p,miR-195-5p,miR-101-3p,miR-216b-5p,miR-16-5p and miR-185-5p expression were assessed by real-time quantitative polymerase chain reaction.The mRNA and protein levels of amyloid precursor protein,amyloid-β40 and amyloid-β42 were evaluated by quantitative real-time polymerase chain reaction,western blot assays and enzyme-linked immunosorbent assays.swAPP695-HEK293 cells were transfected with miR-15b-5p mimic,or treated with 1μM curcumin 24 hours before miR-15b-5p inhibitor transfection.The effects of curcumin on amyloid precursor protein,amyloid-β40 and amyloid-β42 levels were evaluated by western blot assays and enzyme-linked immunosorbent assay.Luciferase assays were used to analyze the interaction between miR-15b-5p and the 3′-untranslated region of amyloid precursor protein.The results show that amyloid precursor protein and amyloid-βexpression were enhanced in swAPP695-HEK293 cells compared with HEK293 parental cells.Curcumin suppressed the expression of amyloid precursor protein and amyloid-βand up-regulated the expression of miR-15b-5p in swAPP695-HEK293 cells.In addition,we found a negative association of miR-15b-5p expression with amyloid precursor protein and amyloid-βlevels in the curcumin-treated cells.Luciferase assays revealed that miR-15b-5p impaired the luciferase activity of the plasmid harboring the 3′-untranslated region of amyloid precursor protein.These findings indicate that curcumin down-regulates the expression of amyloid precursor protein and amyloid-βin swAPP695-HEK293 cells,which was partially mediated by miR-15b-5p via targeting of the 3′-untranslated region of amyloid precursor protein.展开更多
Lateral roots play essential roles in drought tolerance in maize(Zea mays L.). However, the genetic basis for the variation in the number of lateral roots in maize remains elusive. Here, we identified a major quantita...Lateral roots play essential roles in drought tolerance in maize(Zea mays L.). However, the genetic basis for the variation in the number of lateral roots in maize remains elusive. Here, we identified a major quantitative trait locus(QTL),q LRT5-1, controlling lateral root number using a recombinant inbred population from a cross between the maize lines Zong3(with many lateral roots) and 87-1(with few lateral roots).Fine-mapping and functional analysis determined that the candidate gene for qLRT5-1,ZmLRT, expresses the primary transcript for the microRNA miR166a. ZmLRT was highly expressed in root tips and lateral root primordia,and knockout and overexpression of ZmLRT increased and decreased lateral root number,respectively. Compared with 87-1, the ZmLRT gene model of Zong3 lacked the second and third exons and contained a 14 bp deletion at the junction between the first exon and intron,which altered the splicing site. In addition,ZmLRT expression was significantly lower in Zong3 than in 87-1, which might be attributed to the insertions of a transposon and over large DNA fragments in the Zong3 ZmLRT promoter region. These mutations decreased the abundance of mature miR166a in Zong3, resulting in increased lateral roots at the seedling stage.Furthermore, miR166a post-transcriptionally repressed five development-related class-Ⅲ homeodomain-leucine zipper genes. Moreover,knockout of ZmLRT enhanced drought tolerance of maize seedlings. Our study furthers our understanding of the genetic basis of lateral root number variation in maize and highlights ZmLRT as a target for improving drought tolerance in maize.展开更多
In order to find evidence of consistent sequence conservation or the base correlation degree in miRNA,some important sites in the sequences of reported miRNA and their precursors(pre-miRNA) were investigated via infor...In order to find evidence of consistent sequence conservation or the base correlation degree in miRNA,some important sites in the sequences of reported miRNA and their precursors(pre-miRNA) were investigated via information entropy analysis.Twelve different groups of sites were obtained from special locations(head,tail) in miRNAs of different sources according to taxonomy(animal,plant and virus) and then analyzed by measuring the single base information redundancy(D1(L)) and the adjacent base related information redundancy(D2(L)).The results showed that D2(L) has more information than D1(L),though D1(L) changes roughly consistently with D2(L) in each group.Viral pre-miRNAs are more conservative than those belonging to animals or plants.In addition,U is dominant in most sites compared with other nucleotides.It was also found that in the middle of several groups,there were sites where miRNAs were cut down from pre-miRNAs by Enzyme Dicer which were significantly conservative.This phenomenon shows that the conservatism is an aspect of the of miRNA and may be involved in the recognition and cutting by the Dicer.Those results provided another perspective for understanding more about the primary structure of pre-miRNA.展开更多
MicroRNAs（miRNAs） are gene regulators involved in numerous diseases including cancer,heart disease,neurological disorders,vascular abnormalities and autoimmune conditions.Although hsa-mir-499 rs3746444 polymorphism ...MicroRNAs（miRNAs） are gene regulators involved in numerous diseases including cancer,heart disease,neurological disorders,vascular abnormalities and autoimmune conditions.Although hsa-mir-499 rs3746444 polymorphism was shown to contribute to the susceptibility of multiple genes to cancer,the data have yielded conflicting results.Therefore,this meta-analysis was performed to provide a comprehensive assessment of potential association between hsa-mir-499 rs3746444 polymorphism and cancer risk.In this meta-analysis,a total of 9 articles regarding 10 eligible case-control studies in English（including 6134 cases and 7141 controls） were analyzed.No significant association between hsa-mir-499 rs3746444 polymorphism and overall cancer risk was demonstrated.However,an increased risk was observed in the subgroup of breast cancer patients（G allele vs A allele：OR = 1.10,95% CI = 1.00-1.20;P heterogeneity = 0.114;I 2 = 53.9%） and population-based studies（G allele vs A allele：OR = 1.12,95% CI = 1.00-1.25;P heterogeneity = 0.062;I 2 = 64.0%）.The findings suggested an association between hsa-mir-499 rs3746444 polymorphism and increased risk to breast cancer.展开更多
文摘Endometrial cancer （EC） is the most common and lethal gynaecological cancer type in Europe and in North America. Frequently EC arises more in the corpus proper and manifests as round, polypoid expansile masses, but it may also originate in the lower uterine segment or spread in endometrium with necrosis and hemorrhage. The analysis was performed using a custom panel containing all DNA sequences loci coding pre-miRNAs and genes related to biogenesis and regulation of sncRNAs in normal and tumor tissues extracted from 6 unrelated patients with endometrial carcinoma. The identified variations were correlated with mature miRNAs differentially expressed in the same normal and tumor endometrial tissues. The comparison analysis confirmed the high degree of cellular and genetic intratumoral heterogeneity with a temporal and spatial miRNA expression distribution in association with genomic variants identified. The classification of specific DNA mutations, onto the loci identified, should be suitable to characterize possible instability genome regions and help classification of tumors to ameliorate the clinical management of patients affected by endometrial carcinoma.
基金supported by the Science and Technology Planning Project of Guangdong Province of China,No.2016A020226022(to HYL)the Medical and Health Technology Project of Guangzhou of China,No.20161A011068(to HYL)the Guangzhou Science Technology and Innovation Commission of China,No.201704020043(to QCG)
文摘Curcumin exerts a neuroprotective effect on Alzheimer’s disease;however,it is not known whether microRNAs are involved in this protective effect.This study was conducted using swAPP695-HEK293 cells as an Alzheimer’s disease cell model.swAPP695-HEK293 cells were treated with 0,0.5,1,2,5,and 10μM curcumin for 24 hours.The changes in miR-15b-5p,miR-19a-3p,miR-195-5p,miR-101-3p,miR-216b-5p,miR-16-5p and miR-185-5p expression were assessed by real-time quantitative polymerase chain reaction.The mRNA and protein levels of amyloid precursor protein,amyloid-β40 and amyloid-β42 were evaluated by quantitative real-time polymerase chain reaction,western blot assays and enzyme-linked immunosorbent assays.swAPP695-HEK293 cells were transfected with miR-15b-5p mimic,or treated with 1μM curcumin 24 hours before miR-15b-5p inhibitor transfection.The effects of curcumin on amyloid precursor protein,amyloid-β40 and amyloid-β42 levels were evaluated by western blot assays and enzyme-linked immunosorbent assay.Luciferase assays were used to analyze the interaction between miR-15b-5p and the 3′-untranslated region of amyloid precursor protein.The results show that amyloid precursor protein and amyloid-βexpression were enhanced in swAPP695-HEK293 cells compared with HEK293 parental cells.Curcumin suppressed the expression of amyloid precursor protein and amyloid-βand up-regulated the expression of miR-15b-5p in swAPP695-HEK293 cells.In addition,we found a negative association of miR-15b-5p expression with amyloid precursor protein and amyloid-βlevels in the curcumin-treated cells.Luciferase assays revealed that miR-15b-5p impaired the luciferase activity of the plasmid harboring the 3′-untranslated region of amyloid precursor protein.These findings indicate that curcumin down-regulates the expression of amyloid precursor protein and amyloid-βin swAPP695-HEK293 cells,which was partially mediated by miR-15b-5p via targeting of the 3′-untranslated region of amyloid precursor protein.
基金financially supported by the National Key Research and Development Program of China (Grant No. 2016YFD0100801)the National Transgenic Key Project of the Ministry of Agriculture of China (Grant No. 2016ZX08009002)the Natural Science Foundation of Shandong Province (Grant No. ZR2018PC017)。
文摘Lateral roots play essential roles in drought tolerance in maize(Zea mays L.). However, the genetic basis for the variation in the number of lateral roots in maize remains elusive. Here, we identified a major quantitative trait locus(QTL),q LRT5-1, controlling lateral root number using a recombinant inbred population from a cross between the maize lines Zong3(with many lateral roots) and 87-1(with few lateral roots).Fine-mapping and functional analysis determined that the candidate gene for qLRT5-1,ZmLRT, expresses the primary transcript for the microRNA miR166a. ZmLRT was highly expressed in root tips and lateral root primordia,and knockout and overexpression of ZmLRT increased and decreased lateral root number,respectively. Compared with 87-1, the ZmLRT gene model of Zong3 lacked the second and third exons and contained a 14 bp deletion at the junction between the first exon and intron,which altered the splicing site. In addition,ZmLRT expression was significantly lower in Zong3 than in 87-1, which might be attributed to the insertions of a transposon and over large DNA fragments in the Zong3 ZmLRT promoter region. These mutations decreased the abundance of mature miR166a in Zong3, resulting in increased lateral roots at the seedling stage.Furthermore, miR166a post-transcriptionally repressed five development-related class-Ⅲ homeodomain-leucine zipper genes. Moreover,knockout of ZmLRT enhanced drought tolerance of maize seedlings. Our study furthers our understanding of the genetic basis of lateral root number variation in maize and highlights ZmLRT as a target for improving drought tolerance in maize.
基金Supported by the National Natural Science Foundation of China (Grant No. 30600384)the Young Outstanding Scholar Foundation of Northwest Science and Technolgy University
文摘In order to find evidence of consistent sequence conservation or the base correlation degree in miRNA,some important sites in the sequences of reported miRNA and their precursors(pre-miRNA) were investigated via information entropy analysis.Twelve different groups of sites were obtained from special locations(head,tail) in miRNAs of different sources according to taxonomy(animal,plant and virus) and then analyzed by measuring the single base information redundancy(D1(L)) and the adjacent base related information redundancy(D2(L)).The results showed that D2(L) has more information than D1(L),though D1(L) changes roughly consistently with D2(L) in each group.Viral pre-miRNAs are more conservative than those belonging to animals or plants.In addition,U is dominant in most sites compared with other nucleotides.It was also found that in the middle of several groups,there were sites where miRNAs were cut down from pre-miRNAs by Enzyme Dicer which were significantly conservative.This phenomenon shows that the conservatism is an aspect of the of miRNA and may be involved in the recognition and cutting by the Dicer.Those results provided another perspective for understanding more about the primary structure of pre-miRNA.
基金supported by China Natural Science Foundation (No.30901534)Jiangsu Province Natural Science Foundation(No. BK2009444)Grant for the 135 Key Medical Project of Jiangsu Province(No.XK201117)
文摘MicroRNAs（miRNAs） are gene regulators involved in numerous diseases including cancer,heart disease,neurological disorders,vascular abnormalities and autoimmune conditions.Although hsa-mir-499 rs3746444 polymorphism was shown to contribute to the susceptibility of multiple genes to cancer,the data have yielded conflicting results.Therefore,this meta-analysis was performed to provide a comprehensive assessment of potential association between hsa-mir-499 rs3746444 polymorphism and cancer risk.In this meta-analysis,a total of 9 articles regarding 10 eligible case-control studies in English（including 6134 cases and 7141 controls） were analyzed.No significant association between hsa-mir-499 rs3746444 polymorphism and overall cancer risk was demonstrated.However,an increased risk was observed in the subgroup of breast cancer patients（G allele vs A allele：OR = 1.10,95% CI = 1.00-1.20;P heterogeneity = 0.114;I 2 = 53.9%） and population-based studies（G allele vs A allele：OR = 1.12,95% CI = 1.00-1.25;P heterogeneity = 0.062;I 2 = 64.0%）.The findings suggested an association between hsa-mir-499 rs3746444 polymorphism and increased risk to breast cancer.
