细胞永生化对线粒体DNA拷贝数的影响  

The influence of immortalizations on the copy number of mitochondrial DNA

作　　者：张彦春[1] 裴佩[2] 夏昌宇[2] 马祎楠[1] 戚豫[2] 

机构地区：[1]首都医科大学附属北京妇产医院,北京100026 [2]北京大学第一医院实验中心,北京100034

出　　处：《中国优生与遗传杂志》2017年第6期5-7,共3页Chinese Journal of Birth Health & Heredity

基　　金：国家自然科学基金(No.81471153)

摘　　要：目的探讨EB病毒转化形成的永生细胞对线粒体DNA突变比例及拷贝数的影响。方法随机选择23位携带线粒体A3243G突变的患者,采集静脉血7ml,其中2ml提取总DNA,为对照组;5ml用EB病毒转化成为永生细胞,提取总DNA,为实验组。采用Taqman探针联合扩增受阻突变体系定量聚合酶链反应(ARMS-q PCR)方法检测线粒体A3243G突变比例和拷贝数。运用配对t检验进行统计分析。结果实验组的线粒体A3243G突变比例与对照组无明显差异;实验组的野生型线粒体DNA拷贝数,突变型线粒体DNA拷贝数和总线粒体DNA拷贝数与对照组比较,均无明显差异。结论 EB病毒转化对线粒体DNA数量上无明显影响,证实了通过EB病毒转化建立的永生细胞株可以长久保存线粒体DNA,为线粒体相关疾病的研究提供充足的实验材料。Objective： To investigate the immortalized cells that formated of EB virus affect the proportion and copy number of mitochondrial DNA. Methods： Randomly selected 23 patients who carry mutations in the mitochondrial A3243G, draw the venous blood 7ml, in which 2ml to extract the total DNA, as a control group; 5ml to extract the lymphocytes, then used EB virus transformed into immortal cell to extract total DNA, as the experimental group. Using Taqman probe amplification refractory mutation system combined quantitative polymerase chain reaction （ARMS-qPCR） method to detect the ratio of mitochondrial A3243G mutation and copy number. Used paired t test methods for statistical analysis. Results： Compared the control group and the experimental group, the proportion of mitochondrial DNA A3243G, wild-type mitochondrial DNA copy number, the mutant mitochondrial DNA copy number and the total mtDNA copy number all had no significant difference. Conclusion： Immortalized cells by EB virus transformation had no significant effect on the copy number of mitochondrial DNA, and this method could establish long-term preservation of mitochondrial DNA, which could provide sufficient material for the study of mitochondrial diseases.

关 键 词：线粒体DNA 永生化 突变比例 拷贝数 A3243G 

分 类 号：R440[医药卫生—诊断学]