多重PCR-SSCP法检测妇科肿瘤患者hMSH2基因突变  

Detection of hMSH2 gene mutations in gynecologic tumor population by multiple PCR-SSCP

作　　者：梁立治[1] 何云[2] 庄志雄[3] 何春华[3] 刘汝青[2] 彭小萍[1] 

机构地区：[1]中山大学肿瘤医院妇科,广东广州510060 [2]中山大学公共卫生学院卫生毒理学教研室,广东广州510080 [3]深圳市疾病预防控制中心,广东深圳518020

出　　处：《中国癌症杂志》2003年第2期128-130,共3页China Oncology

基　　金：国家自然科学基金项目 (No .30 2 0 0 2 30 );国家重点基础研究发展规划项目 ( 973计划 ) (No .2 0 0 2CB512 90 3;2 0 0 2CB512 90 4 );广东省自然科学基金项目 (No .0 2 1816 )

摘　　要：目的 :建立经济快捷地筛查基因突变的技术 ,研究妇科肿瘤患者hMSH2基因突变情况 ,寻找与妇科肿瘤发生相关的分子标记物。方法 :通过PCR优化策略 ,建立多重PCR SSCP法 ;收集 42名妇科肿瘤病人的基础资料及血液标本 ,抽提血液DNA ,用多重PCR法扩增hMSH2基因第六、第七外显子 ,并进行单链构象多态性分析(SSCP)及DNA序列分析。结果 :检测出 2位妇科肿瘤病人在hMSH2基因第七外显子有杂合性突变 ,且为Leu→Phe的错义突变。未发现hMSH2基因第六外显子上存在突变。结论 :多重PCR SSCP是筛查基因突变的一种快速有效的方法 ,hMSH2基因第七外显子上的突变可能是与妇科肿瘤发生相关的一种分子标记物。Purpose:To set up a new method to screen hMSH2 gene mutations, to detect hMSH2 gene mutations in gynecologic tumor population and to find molecular biomarkers of tumor. Methods:The basic data and blood samples from 42 gynecologic tumor patients werre collected. The genetic mutations of the sixth exon and seventh exon of hMSH2 gene were investigated by multiple polymerase chain reaction single strand conformation polymorphism (PCR SSCP),followed by DNA sequencing. Results:The successes in setting up the multiple PCR SSCP method made it possible to detect hMSH2 gene mutations more quickly and more economically. Two samples of the seventh exon mutation was detected in the tumor population. The mutation is Leu→Phe missense mutation. No mutation of the sixth exon was detected by SSCP.Conclusions:The multiple PCR SSCP method is effective in detecting genetic mutations. There is a mutation site in the seventh exon of hMSH2 gene. It is probably a biomarker of gynecologic tumor. This discovery might offer the basis for further investigation of mutations in large amount population and studies of the function of the mutation.

关 键 词：多重PCR-SSCP法 检测 妇科患者 HMSH2 基因突变 

分 类 号：R711[医药卫生—妇产科学]