变性高效液相色谱-测序技术在检测乳腺癌BRCA1基因突变中的临床诊断价值  

The clinical value of denaturing high performance liquid chromatography in detecting mutations of breast cancer susceptibility gene-1

作　　者：李丹[1] 吴永和[1] 闵大六[2] 汪朝晖[1] 郑俊峰[1] 孙大光[1] 马旭[1] 

机构地区：[1]国家人口计划生育委科学技术研究所遗传研究室中国医学科学院中国协和医科大学,北京100081 [2]徽省肿瘤医院

出　　处：《中华检验医学杂志》2006年第7期643-646,共4页Chinese Journal of Laboratory Medicine

基　　金：国家人类遗传资源平台计划资助项目(2004DKA30490)

摘　　要：目的探讨变性高效液相色谱(DHPLC)-测序技术在早期检测乳腺癌BRCA1基因突变的临床意义。方法用聚合酶链反应(PCR)扩增BRCA1基因,该扩增产物经琼脂糖凝胶电泳鉴定后,直接进行DHPLC分析;有峰型改变的样本作测序分析,以确认突变。结果从124例乳腺癌患者中共发现9种突变。包括7种错义突变,其中,1例Stop598,1例A807E合并E809L,1例A807E,2例Y856H,1例T967S,2例I1170F,1例R1203G;一种存在于一号内含子的突变,即IVS101-10 T>C 17例;一种2号外显子的5’非翻译区突变,即A118T 2例。此外,在乳腺癌标本和正常对照标本中都发现有几处常见的单核苷酸多态性位点。结论本研究确定了9种乳腺癌患者可能发生的突变类型,为今后临床应用DHPLC对高危人群BRCA1基因进行早期基因检测提供了9个位点。DHPLC-测序技术可快速、方便地区分带有碱基替换和小片段缺失的DNA片段,是一种可用于早期临床诊断乳腺癌BRCA1基因改变的高效、灵敏和操作简便的方法。Objective To estimate the clinical value of DHPLC-Sequencing in detecting BRCA1 gene mutations in initial stages of breast cancer and establish a rapid and efficient method for the use in clinical laboratory. Methods BRCA1 gene was amplified the PCR products were identified by the agarose gel electrophoresis and analyzed by DHPLC. The samples with abnormal profiles were sequenced and judged whether to contain mutations or not. Results 124 Chinese breast cancer patients were enrolled in the study. Nine types of mutations were identified. Among them, there were seven missense：Stop598 （one case）, A807E＆E809L（ one case） ,A807E （ one case） ,Y856H （ twe case） ,T967S（ one case） ,I1170F（ two cases） . R1203G（one case） and one IVS101 -10T 〉 C point-mutation （seventeen cases） and one point-mutation at A118T in the 5＇ UTR of Exon2（ two cases）. Besides, several kinds of SNPs were found both in breast cancer samples and in normal contrast samples. Conclusions We identified 9 types of mutation that early-onset breast cancer patients might have could be used as destination sites during clinical detection of BRCA1 gene mutations. It＇ s easy to seek out the samples with substitution, deletion or insertion mutations by the use of DHPLC-Sequencing. The method of DHPLC-Sequencing is an efficient, sensitive and rapid method for the clinical molecular diagnosis for gene mutation.

关 键 词：变性高效液相色谱 乳腺肿瘤 基因突变 

分 类 号：R737.9[医药卫生—肿瘤]