AIM:To investigate the reciprocal modulation between microRNA(miRNA) and DNA methylation via exploring the correlation between miR-373 and methyl-CpGbinding domain protein(MBD)2.METHODS:MiR-373 expression was examined...AIM:To investigate the reciprocal modulation between microRNA(miRNA) and DNA methylation via exploring the correlation between miR-373 and methyl-CpGbinding domain protein(MBD)2.METHODS:MiR-373 expression was examined using the TaqMan miRNA assay.Methylation of miR-373 was investigated using methylation-specific polymerase chain reaction,and recruitment of methyl binding proteins was studied using the chromatin immunoprecipitation assay.Mutation analysis was conducted using the QuikChange Site-Directed Mutagenesis kit.The activity of miR-373 gene promoter constructs and targeting at MBD2-three prime untranslated region(3'UTR) by miR-373 were evaluated by a dual-luciferase reporter gene assay.RESULTS:In hilar cholangiocarcinoma,miR-373 decreased and was closely associated with poor cell differentiation,advanced clinical stage,and shorter survival.The promoter-associated CpG island of miR-373 gene was hypermethylated and inhibited expression of miR-373.MBD2 was up-regulated and enriched at the promoter-associated CpG island of miR-373.Methylation-mediated suppression of miR-373 required MBD2 enrichment at the promoter-associated CpG island,and miR-373 negatively regulated MBD2 expression through targeting the 3'UTR.CONCLUSION:MiR-373 behaves as a direct transcriptional target and negative regulator of MBD2 activity through a feedback loop of CpG island methylation.展开更多
目的分析microRNA-373(miR-373)及直接靶标基因TRPS1在喉癌组织中的表达水平及其临床意义。方法收集2018年1月~2019年12月锦州医科大学附属第一医院耳鼻咽喉头颈外科确诊为喉癌并行手术治疗的55例患者的喉癌组织及42例癌旁正常上皮组织...目的分析microRNA-373(miR-373)及直接靶标基因TRPS1在喉癌组织中的表达水平及其临床意义。方法收集2018年1月~2019年12月锦州医科大学附属第一医院耳鼻咽喉头颈外科确诊为喉癌并行手术治疗的55例患者的喉癌组织及42例癌旁正常上皮组织。所有标本在取材前均未行任何抗肿瘤治疗,采用SYBR Green qRT-PCR方法检测患者癌组织及对应癌旁正常上皮组织中miR-373表达情况,采用免疫荧光法测量其靶标基因TRPS1在喉癌组织中的表达水平,并分析miR-373、TRPS1与临床病理特征的关系。结果miR-373及TRPS1在喉癌组织中的表达均高于癌旁正常上皮组织(P<0.05);不同TNM分期和淋巴结转移间miR-373相对表达量及TRPS1比较,差异有统计学意义(P<0.05);但不同年龄、性别、吸烟、饮酒及分化程度间miR-373相对表达量及TRPS1比较,差异无统计学意义(P>0.05)。结论miR-373及TRPS1在喉癌组织中的表达升高,且与淋巴结转移及肿瘤TNM分期有关。展开更多
基金Supported by National Natural Science Foundation of China,No. 81071998Hubei Natural Science Foundation,No.2008CDB159Specialized Research Fund for the Doctoral Program of Higher Education,No. 20070487114
文摘AIM:To investigate the reciprocal modulation between microRNA(miRNA) and DNA methylation via exploring the correlation between miR-373 and methyl-CpGbinding domain protein(MBD)2.METHODS:MiR-373 expression was examined using the TaqMan miRNA assay.Methylation of miR-373 was investigated using methylation-specific polymerase chain reaction,and recruitment of methyl binding proteins was studied using the chromatin immunoprecipitation assay.Mutation analysis was conducted using the QuikChange Site-Directed Mutagenesis kit.The activity of miR-373 gene promoter constructs and targeting at MBD2-three prime untranslated region(3'UTR) by miR-373 were evaluated by a dual-luciferase reporter gene assay.RESULTS:In hilar cholangiocarcinoma,miR-373 decreased and was closely associated with poor cell differentiation,advanced clinical stage,and shorter survival.The promoter-associated CpG island of miR-373 gene was hypermethylated and inhibited expression of miR-373.MBD2 was up-regulated and enriched at the promoter-associated CpG island of miR-373.Methylation-mediated suppression of miR-373 required MBD2 enrichment at the promoter-associated CpG island,and miR-373 negatively regulated MBD2 expression through targeting the 3'UTR.CONCLUSION:MiR-373 behaves as a direct transcriptional target and negative regulator of MBD2 activity through a feedback loop of CpG island methylation.
文摘目的分析microRNA-373(miR-373)及直接靶标基因TRPS1在喉癌组织中的表达水平及其临床意义。方法收集2018年1月~2019年12月锦州医科大学附属第一医院耳鼻咽喉头颈外科确诊为喉癌并行手术治疗的55例患者的喉癌组织及42例癌旁正常上皮组织。所有标本在取材前均未行任何抗肿瘤治疗,采用SYBR Green qRT-PCR方法检测患者癌组织及对应癌旁正常上皮组织中miR-373表达情况,采用免疫荧光法测量其靶标基因TRPS1在喉癌组织中的表达水平,并分析miR-373、TRPS1与临床病理特征的关系。结果miR-373及TRPS1在喉癌组织中的表达均高于癌旁正常上皮组织(P<0.05);不同TNM分期和淋巴结转移间miR-373相对表达量及TRPS1比较,差异有统计学意义(P<0.05);但不同年龄、性别、吸烟、饮酒及分化程度间miR-373相对表达量及TRPS1比较,差异无统计学意义(P>0.05)。结论miR-373及TRPS1在喉癌组织中的表达升高,且与淋巴结转移及肿瘤TNM分期有关。
