变性高效液相色谱检测乳腺癌患者血浆线粒体D环HVR2突变  

DHPLC detection of mtDNA D-loop HVR2 gene mutation in plasma DNA in patients with primary breast cancer

作　　者：靳斌[1] 任军[1] 张亮[1] 汪海丹[1] 白俊[1] 张燕军[1] 斯小明[1] 潘伯荣[1] 

机构地区：[1]第四军医大学西京医院肿瘤中心,陕西西安710033

出　　处：《第四军医大学学报》2003年第10期911-913,共3页Journal of the Fourth Military Medical University

基　　金：全国高校优秀青年教师教学科研奖励计划资助(TRAPOYT 99 1 6);高等学校骨干教师资助计划 (FUKT2 0 0 0 0 6)资助项目

摘　　要：目的 :探讨扩增乳腺癌患者血浆线粒体DNA(mito chondrialDNA ,mtDNA)的可行性 ,研究乳腺癌患者血浆线粒体DNA基因突变检测对于乳腺癌早期诊断及其预后的意义 .方法 :选取乳腺癌患者 10例 ,正常健康者 10人 ,提取血浆DNA ,采用聚合酶链反应 (PCR)扩增血浆线粒体DNA的D环HVR2 ,用两对引物扩增 ,用变性高效液相色谱法 (DHPLC)对产物进行突变筛选 .结果 :10例乳腺癌患者血浆提取的DNA能扩增出 11个目的片断 ,DHPLC检测到有 4例突变 ,正常健康者 10人扩增出 1例 .结论 :乳腺癌患者血浆DNA扩增与正常组有明显差异 (P <0 .0 5 ) 。AIM: To evaluate the presence of mtDNA in plasma from primary breast cancer patients and to study the clinical significance of mtDNA D Loop gene mutations in early diagnosis and prognosis of breast cancer. METHODS: The plasma DNA in 10 patients with primary breast cancer and 10 normal healthy people was extracted and mtDNA D loop HVR2 fragments were amplified with two pairs of primers by PCR. The mutation of production was screened by denaturing high performance liquid chromatography (DHPLC). RESULTS: Eleven target fragments were obtained from the 10 primary breast cancer. Four fragments of mutation were detected by DHPLC. One target fragment was obtained from the healthy control. CONCLUSION: Plasma DNA in primary breast cancer is different from that in healthy people. MtDNA D loop gene mutations are of significance in the early diagnosis and prognosis in breast cancer.

关 键 词：乳腺肿瘤 DNA 线粒体 高变区2 D环 血浆 色谱法 液相 

分 类 号：R737.9[医药卫生—肿瘤]