荧光实时定量PCR检测非小细胞肺癌组织及外周血中RRM1 mRNA表达水平的方法  

Detection of expression of RRM1 mRNA by real-time fluorescent quantitative PCR in non-small cell lung cancer tissues and peripheral blood

作　　者：董嵩[1] 吴一龙[2] 郭爱林[1] 陈志红[1] 谢至[1] 林嘉颖[1] 张绪超[1] 

机构地区：[1]广东省人民医院医学研究中心生物芯片部,广州510080 [2]广东省人民医院肿瘤中心,广州510080

出　　处：《肿瘤》2007年第7期577-581,共5页Tumor

基　　金：广州市科技局重点科技攻关计划资助(编号:2001-Z-044-01);广东省重点医学科技攻关专项课题资助(编号:2003-1);广东省名医工程资助(编号:2004-25)

摘　　要：目的:建立检测非小细胞肺癌患者肿瘤组织及外周血RRM1 mRNA表达水平的实时定量荧光PCR方法,比较不同标本的检测方法及效果,为临床应用该基因表达水平预测药物疗效提供方法基础。方法:构建RRM1基因的质粒标准品,运用ABI7000PCR仪进行实时定量分析,探索最佳反应条件并建立标准曲线用以检测临床标本。结果:检测了18例患者的肺癌及正常肺组织标本,同时检测了17例患者的外周血标本,以β-actin为管家基因计算得肺癌组织相对表达量平均为4.46×10-3,正常肺组织为3.43×10-3,外周血为2.54×10-3。Ct值与模板起始浓度对数值之间有良好的线性相关。结论:所建SYBRGreenⅠ荧光定量PCR方法能成功检测肺癌组织及外周血单个核细胞RRM1 mRNA表达水平,检测方法基本相同,有较高的敏感性与特异性,可应用于临床检验工作。Objective： We tired to develop a method of detecting the expression level of RRM1 mRNA in tumor tissues and peripheral blood by real-time fluorescent quantitative PCR and compare the detection methods and results between the two different samples. We aimed to provide the basis for predicting clinical outcome from the expression of RRMI gene. Methods：The plasmid standard of RRM1 gene was constructed. Real-time quantitative analysis was performed using ABI7000 PCR kit. The PCR condition was optimized and the standard curve was established to detect the clinical samples. Results： The RRMI gene expression was detected in lung cancer tissues and normal lung tissues from 18 patients and in peripheral blood samples from 17 patients. The relative expression of RRM1 normalized by β-actin was 4.46×10^-3 in lung cancer tissues, 3.43×10^-3 in normal tissues, 2.54×10^-3 in peripheral blood. The linear correlation between Ct value and the logarithm of original concentration was favorable. Conclusion： The expression of RRM1 mRNA in non-small cell lung cancer tissues and peripheral blood was successfully detected by using SYBR Green Ⅰ fluorescence realtime quantitative PCR assay. There is no significant difference between the two sources of samples. The detection method has relatively higher sensitivity and specificity and can be used in clinical analysis.

关 键 词：癌 非小细胞肺 核苷酸还原酶类 基因表达 实时定量荧光PCR 

分 类 号：R730.43[医药卫生—肿瘤] R734.2[医药卫生—临床医学]