外周血基因表达分析内参基因的优选  

Determination of suitable reference genes for gene expression studies in human peripheral blood

作　　者：宋莉[1] 李风华[2] 孙莹莹 宋卫华[1] 宋燕[1] 柳婷婷 田雪晴[1] 肖宁[1] 温红艳 张银辉[1] 孟宪敏[1] 陈敬洲[1] SONG Li;LI Feng-hua;SUN Ying-ying;SONG Wei-hua;SONG Yan;LIU Ting-ting;TIAN Xue-qing;XIAO Ning;WEN Hong-yan;ZHANG Yin-hui;MENG Xian-min;CHEN Jing-zhou(State Key Laboratory of Cardiovascular Disease ,Fuwai Hospital,National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College ,Beijing 100037, China;：Department of Internal Medi- cine,Peking University Hospital,Beijing 100871 ,China)

机构地区：[1]中国医学科学院,北京协和医学院,国家心血管病中心,阜外医院,心血管疾病国家重点实验室,北京100037 [2]北京大学医院内科,北京100871

出　　处：《实用医院临床杂志》2018年第3期5-8,共4页Practical Journal of Clinical Medicine

基　　金：中央高校基本科研业务费专项资金资助项目(编号:3332015173);国家自然科学基金资助项目(编号:30940029)

摘　　要：目的明确外周血RNA定量分析最佳内参基因组合。方法采用实时荧光定量PCR方法检测5种管家基因的表达情况,应用qBASE+软件对基因表达稳定性进行分析并筛选最佳内参基因组合。结果采用冠心病病例对照人群共98例,常用内参基因ACTB的表达水平变化最大。内参基因的表达稳定度由高到低依次为DECR1>GAPDH>TRAP1>PPIB>ACTB,并分析出最佳内参基因组合为DECR1、GAPDH和TRAP1。结论研究结果为冠心病外周血的内参基因的选择提供了可靠的依据,可为该类样品的基因表达定量分析在基础研究和临床诊断的应用提供技术支撑。Objective To identify the optimal set of the most stably expressed gene in human peripheral blood. Methods Quantitative real-time PCR was used to determine the transcription profiles of 5 housekeeping genes, qBASE＋ software was used to ana- lyze the gene＇s stabilities and choose the optimal set of reference genes. Results The total 98 case and control samples are collected from patients with coronary heart disease. The Ct range of ACTB, as the most eommonly used reference gene,was ehanged widely from 20. 09 （ 30. 06. The stability of these genes ranked in the following order： DECR1 〉 GAPDH 〉 TRAP1 〉 PPIB 〉ACTB, and the optimal reference genes set was composed of DECR1, GAPDH, and TRAP1. Conclusion The results supply a reliable evidence for choosing the reference genes in human peripheral blood, and provide a teehnieal support for application of gene expression analysis in the basic researeh and clinic diagnosis.

关 键 词：实时定量PCR 内参基因 qBASE+软件 外周血 

分 类 号：R34[医药卫生—基础医学]