变性高效液相色谱法筛选XRCC4基因突变  

Screening Gene Mutations of XRCC4 by Denaturing High-performance Liquid Chromatography

作　　者：贾静[1] 潘建基[1] 苏颖[1] 陈增[1] 邹长棪[1] 陈传本[1] 潘才柱[1] 

机构地区：[1]福建医科大学教学医院,福建省肿瘤医院放射生物研究室,福州350014

出　　处：《福建医科大学学报》2009年第3期241-243,共3页Journal of Fujian Medical University

基　　金：福建省医学创新课题(2001-CX-15)

摘　　要：目的应用变性高效液相色谱技术(DHPLC)-异源双链分析对鼻咽癌患者XRCC4基因突变进行筛查与鉴定,研究鼻咽癌患者XRCC4的基因突变情况,探讨DHPLC在筛查相关基因突变、预测放疗敏感性方面的应用。方法采用聚合酶链反应(PCR)和DHPLC筛查88例鼻咽癌患者XRCC4基因的突变。结果对DHPLC图形异常的PCR扩增片断进行DNA测序鉴定突变位点及性质,检测出28例XRCC4基因8号外显子第377位C变成T,导致126号密码子的ser→phe,导致氨基酸残基的替代变化(丝氨酸→苯丙氨酸)。结论用PCR-DHPLC筛查结合测序分析检测XRCC4突变是一种高效、经济、简便、可靠的方法。Objective To detect mutations in XRCC4 gene in Nasopharyngeal carcinoma （NPC） patients by denaturing high-perfozmance liquid chromatography （DHPLC）and to investigate the potential application of DHPLC in screening gene mutation of XRCC4 for prediction of radio therapy sensitivity. Methods polymerase chain reaction and denaturing high-performance liquid chromatography （DHPLC） were used to screen XRCC4 mutations in 88 patients of NPC. Results The PCR-amplified XRCC4 DNA fragments with a different DHPLC elution profile from their corresponding wild-type profile were sequenced to identify the position and type of mutation. A mutation in the exon8 （377 C→T） were found in 28 Patients, which leads to an amino-acid substitution （serine→phenylalanine）. Conclusions PCR-DH- PLC is an effective, economical, and simple method with reliability for detecting mutations in XRCC4 gene.

关 键 词：鼻咽肿瘤 辐射耐受性 突变 肿瘤细胞 培养的 DNA修复 色谱法 高压液相 基因型 

分 类 号：R739.630[医药卫生—肿瘤] R394.2[医药卫生—临床医学]