采用半定量RT-PCR方法检测20例单纯性马蹄内翻足患儿下肢肌肉及肌腱组织中COL1A1基因mRNA的表达, 根据COL1A1基因转录调控区-1 031 bp~ +30 bp及第1内含子的序列, 设计8对引物, PCR扩增后, 采用变性梯度凝胶电泳技术筛查突变并测序。半定量RT-PCR结果表明, 与正常对照组相比, 单

HEREDITAS (Beijing)

2008年6月, 30(6): 723―727 ISSN 0253-9772 www.wendangwang.com

研究报告

DOI: 10.3724/SP.J.1005.2008.00723

COL1A1基因转录调控序列变异与单纯性马蹄内翻足的相关性

赵宁, 金春莲, 刘丽英, 曹东华, 林长坤, 吉士俊, 孙开来

中国医科大学医学遗传学教研室, 沈阳 110001

摘要: 采用半定量RT-PCR方法检测20例单纯性马蹄内翻足患儿下肢肌肉及肌腱组织中COL1A1基因mRNA的表达, 根据COL1A1基因转录调控区 1 031 bp~ +30 bp及第1内含子的序列, 设计8对引物, PCR扩增后, 采

用变性梯度凝胶电泳技术筛查突变并测序。半定量RT-PCR结果表明, 与正常对照组相比, 单纯性马蹄内翻足患儿患侧肌肉及肌腱组织中COL1A1基因表达水平明显上调(t =12.680, P＜0.05); 经PCR-DGGE筛查并测序发现1名患者存在 161(C→T)的杂合变异, 另1名患者存在+274(C→G)的纯合变异。二者均为新发现的变异。提示COL1A1基因转录调控序列变异可能是单纯性马蹄内翻足的致病原因之一。

关键词: 单纯性马蹄内翻足; COL1A1基因; 反转录聚合酶链反应; 基因变异; 变性梯度凝胶电泳

Association study between mutations of transcription regulator sequences of COL1A1 gene and idiopathic con-genital talipes equinovarus

ZHAO Ning, JIN Chun-Lian, LIU Li-Ying, CAO Dong-Hua, LIN Chang-Kun, JI Shi-Jun, SUN Kai-Lai

Department of Medical Genetics, China Medical University, Shenyang 110001, China

Abstract: RT-PCR was used to detect the expressions of COL1A1 mRNA in 20 patients with idiopathic congenital talipes equinovarus (ICTEV). The primers were designed by Primer 5 according to sequences of 1 031 bp~+30 bp and the first

intron of COL1A1. PCR-DGGE was used to screen the mutations in COL1A1 gene. Expression of COL1A1 on mRNA lev-els showed significantly higher in patients with ICTEV than in normal persons (t=12.680, P<0.05). By DNA sequencing, a

161(T→C) heterozygous mutation and a+ 274(C→G) homozygous mutation were detected, and both were new identified

mutations. These results indicated that the mutations in transcription regulator sequences of COL1A1 could cause ICTEV.

Keywords: idiopathic congenital talipes equinovarus; COL1A1 gene; reverse transcription-polymerase chain reaction; gene

mutation; denaturing gradient gel electrophoresis

单纯性马蹄内翻足(idiopathic congenital talipes equinovarus, ICTEV)是一种常见的严重危害儿童健

康的先天性足部畸形。其发病率与种族及性别有明显关系, 在我国约为0.6‰~1‰, 男女之比约为2.6︰1,

收稿日期: 2007 10 29; 修回日期: 2008 02 20

基金项目: 国家自然科学基金(编号: 30471803)和国家重点基础研究发展规划项目(编号: 2001CB510301)资助[Supported by the National Natural Science Foundation of China (No. 30471803) and the National Basic Research Program (No. 2001CB510301)]

作者简介: 赵宁(1978 ), 女, 辽宁人, 硕士研究生, 研究方向：遗传病的分子遗传学。Tel: 13130220918; E-mail: ningzhao78@hotmail.com 通讯作者: 金春莲(1945 ), 女, 朝鲜族, 辽宁人, 教授, 研究方向：遗传病的分子遗传学。Tel: 024-23256666-5324; E-mail: chunlianjin@126.com