文章摘要
李伟,胡宝春,龚 照.广东汉族妊娠期糖尿病妇女胎盘组织microRNA差异表达谱分析[J].南京医科大学学报,2017,(3):298~302
广东汉族妊娠期糖尿病妇女胎盘组织microRNA差异表达谱分析
Investigation of microRNA expression profile of placental tissues between gestational diabetes mellitus(GDM)and normal pregnant women
投稿时间:2015-09-14  
DOI:10.7655/NYDXBNS20170308
中文关键词: 妊娠期糖尿病  miRNA  差异表达谱  高通量测序
英文关键词: gestational diabetes mellitus  miRNA  expression profile  high through-put sequencing
基金项目:广东省医学科研基金(A2013452)
作者单位
李伟 武警广东酋总队医院内分泌科 
胡宝春 武警广东酋总队医院内分泌科 
龚 照 妇产科广东广州510507 
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中文摘要:
      目的:探索妊娠期糖尿病(gestational diabetes mellitus,GDM)与正常妊娠胎盘组织微小RNA(microRNA,miRNA)的差异表达,为深入研究GDM发病机制提供基础。方法:选取于2013年1月-2014年1月在武警广东省总队医院、南方医科大学珠江医院就诊并分娩的GDM孕妇20例作为观察组(GDM组),均为粤籍汉族并且相互之间无亲缘关系,随机选取同期无妊娠合并症的正常广东汉族孕妇20例作为对照组。胎儿分娩岳收集胎盘组织,用TRIzol法进行总RNA提取,经过质量和纯度分析鉴定合格岳分别选取5个样本混合成GDM和对照组,采用逆转录法构建小RNA文库,使用第二代高通量测序采用边合成边测序的方法对miRNA进行测序并分析结果。结果:从胎盘组织提取到的总RNA纯度较高,D(260 nm)/D(280 nnl)>1.90,并成功构建小RNA的cDNA文库。采用高通量测序得到长度为10~35 nt的小RNA片段,其中文库数量最多的小RNA长度21~24 nt,miRNA量占所有小RNA总量的30%以上,最终获得洁净的片段超过10 000 000。将得到的片段与Genebank及Rfram数据库比对,去除其他小RNA筛选出miRNA达7 000多种。通过样本两两比较分析,与对照组相比,发现GDM组有138种已知的miRNA表达明显上调,包括has-miR-548au-5p、has-miR-95a-5p、has-miR-373-5p、has-miR-216-5p等(10g2-ratio>l,P<0.01);16种已知miRNAs明显表达下调,如has-miR-5699-5p、has-miR-4286等(10g2-ratio<-1,P<0.01)。Mireap软件预测出27种新miRNA,其中有9种在GDM表达升高,6种表达降低。结论:GDM的胎盘组织miRNAs与对照组存在表达差异,胎盘组织miRNA可能在GDM发病过程中起一定的调节作用。
英文摘要:
      Objective:To investigate microRNA(miRNA)expression profile of placental tissues between gestational diabetes mellitus (GDM)and normal pregnant women.Methods:A total of 20 cases of pregnant women with GDM were randomly selected from Armed Police Hospital Affiliated to Guangzhou Medical College and Zhujiang Hospital of Southern Medical University in January 2013 to January 2014,and notmal pregnant Han women from Guangdong without complications in the same period were enrolled as control.Placenta was obtained after delivery tollowed by total RNA extraction with Trizol.Five samples of total RNA were randomly picked and mixed as the GDM and notmal control group,respectively,for next step.miRNA library of GDM and control was built by the reverse transcription method and used for investigation of expression of various miRNAs by high throughput sequencing.Results:Purified total RNA was extracted from placenta and miRNA library was successfull constructed.After transition from raw data,there were over 10 000 000 clean reads with 21-24 nt-long small RNA the most among which miRNAs were over 30% in each group.After removal of other RNA by comparison between Rfam and Genebank database over 7 000 miRNA species were accounted for.A total of
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