唐氏综合征胎儿21号染色体上新微小RNA基因的鉴定和功能分析

J4 ›› 2012, Vol. 38 ›› Issue (6): 1141-1146.

唐氏综合征胎儿21号染色体上新微小RNA基因的鉴定和功能分析

李武县¹|徐勇^1,2|刘雪燕^3,秦晓林¹|梁勤东¹|戴勇^1,4|涂植光^1

1. 重庆医科大学教育部临床检验诊断学重点实验室|重庆 400016；2.广东省深圳市坪山新区人民医院|广东深圳518118；3. 广东省深圳市人民医院急诊ICU|广东深圳518020；4.广东省深圳市人民医院临床医学研究中心|广东深圳 518020

收稿日期:2012-02-06 发布日期:2012-11-28
通讯作者: 涂植光（Tel：023-68485797，E-mail: tuzhiguang@yahoo.com.cn） E-mail:tuzhiguang@yahoo.com.cn
作者简介:李武县（1985-）|男|重庆市人|在读医学硕士|主要从事分子遗传学与生物化学方面的研究。 
基金资助:
广东省科技厅科技计划项目资助课题(2012B032000008)；深圳市重点科技计划资助课题（201001006）

Identification and functional analysis of chromosome 21-derived new microRNA gene in Down syndrome fetuses

LI Wu-xian1,XU Yong^1,2,LIU Xue-yan³,QIN Xiao-lin_¹,LING Qin-dong¹,DAI Yong^1,4,TU Zhi-guang¹

1. Key Laboratory of Clinical Medical Diagnostics,Ministry of Education,Chongqiong Medical University,Chongqing 400016,China|2. Pingshan People’s Hospital,Guangdong Province,Shenzhen 518118,China;3. Emergency Intensive Care Unit,Shenzhen People’s Hospital,Guangcdong Province,Shenzhen 518020,China|4. Clinical Medical Research Center,Shenzhen People’s Hospital,Guangdong Province,Shenzhen 518020,China）

Received:2012-02-06 Published:2012-11-28

摘要/Abstract

摘要：

目的：利用Solexa高通量深度测序技术鉴定21号染色体上新的微小RNA(miRNA)基因，为阐明21号染色体功能、探索唐氏综合征(DS)患者新的治疗靶点提供依据。方法：收集5例经染色体核型分析诊断为DS胎儿的脐带血和3例正常胎儿脐带血，分析染色体核型；提取胎儿脐带血单个核细胞总RNA，构建小RNA文库，采用Solexa高通量深度测序、计算机分析、Stem-loop RT-PCR法鉴定 miRNA基因并分析其生物学功能。结果:血染色体核型分析，5例DS胎儿脐带血显示DS标准核型，3例正常胎儿脐带血显示正常核型；在DS胎儿脐带血中共发现21个新miRNA，其中仅1个候选miRNA来源于21号染色体，位于21号染色体的“DS关键区域”，命名为“miR-nov21”，后者在DS胎儿脐带血单个核细胞中表达水平高于正常胎儿。生物学功能分析，miR-nov21可调节211个靶基因共215个位点，与基因的表达、细胞分化的调节、胚胎形态的发生和心脏的发育有关。结论: 21号染色体“DS关键区域”存在1个新的miRNA基因——miR-nov21。

关键词: 微小RNA；唐氏综合征；高通量深度测序技术；脐带血

Abstract:

Objective To identify the novel microRNA(miRNA) gene located at the chromosome 21 by Solexa high-throughput sequencing technology,and to provide basis for clarifying function of the chromosome 21 and investingating the new therapeutic target of Down syndrome(DS) patients.Methods Thecord blood of 5 DS fetues verified by chromosome karyotype analysis and 3 normal fetuses were collected.Total RNA was extracted from mononuclear cells of cord blood in DS fetuses to construct the small RNA library.Then the miRNA gene was identified by Solexa high-throughput sequencing,computer analysis and Stem-loop RT-PCR,and its biological function was analyzed.Results The chromosome karyotype analysis showed that the karyetypes in cord blood of DS fetuses were standard DS karyotypes,and those in normal fetuses were normal karyotypes. A total of 21 novel miRNAs were identified in cord blood of DS fetuses.But only one candidat miRNA was located on the “DS critical region” of the chromosome 21,named as “miRnov21”, and its expression level in mononuclear cells of cord bood in DS fetuses was higher than that in normal fetuses.Biological funcation analysis revealed that miR-nov21 could regulate 211 conserved target genes containing 215 conserved sites,which were involved in gene expression,regulation of cell differentiation,embryonic morphogenesis,and heart development.Conclusion There is a novel miRNA gene in “DS critical region” of the chromosome 21.

Key words: microRNA, Down syndrome, high-throughput sequence technology, cord blood

中图分类号:

R596

李武县|徐勇|刘雪燕,秦晓林|梁勤东|戴勇|涂植光. 唐氏综合征胎儿21号染色体上新微小RNA基因的鉴定和功能分析[J]. J4, 2012, 38(6): 1141-1146.

LI Wu-xian,XU Yong,LIU Xue-yan,QIN Xiao-lin,LING Qin-dong,DAI Yong. Identification and functional analysis of chromosome 21-derived new microRNA gene in Down syndrome fetuses[J]. J4, 2012, 38(6): 1141-1146.