Choice of endogenous control for gene expression in nonsmall cell lung cancer
Choice of endogenous control for gene expression in nonsmall cell lung cancer
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DOI：
10.1183/09031936.05.00050205
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发表时间：
2005-12-01
影响因子：
24.3
通讯作者：
Liu, HP
中科院分区：
文献类型：
--
作者：
Liu, DW;Chen, ST;Liu, HP
本研究旨在寻找一种适合于非小细胞肺癌（NSCLC）组织中实时荧光定量PCR检测的内源性调控基因。（甘油醛-3-磷酸脱氢酶（GAPDH）、v-abl Abelson鼠白血病病毒癌基因同源物1、β-2-微球蛋白、次黄质磷酸核糖基转移酶1（HPRT 1）、磷酸甘油酸激酶1、肽基脯氨酰异构酶A和核糖体蛋白，large，P0）在18例异质性NSCLC肿瘤标本、10例正常肺组织和6个NSCLC细胞系中进行了定量RTPCR分析。比较各对照基因的循环阈值（Ct）在3个组织组和亚组中的方差和相关系数。计算每个对照基因的Ct值与其余对照基因的平均Ct值之间的差异和相关系数，GAPDH基因转录本显示出最小的方差，线性回归分析表明GAPDH和HPRT在合并的肿瘤和正常肺组织中具有最强的相关性。GAPDH的表达量与其他内源性对照基因的平均表达量之间的相关性最小，在7个常见的内源性对照基因中，3-磷酸甘油醛脱氢酶最适合用于非小细胞肺癌组织标本的定量RT-PCR反应。
This study attempts to identify a suitable endogenous control gene for real-time RTPCR in nonsmall cell lung cancer (NSCLC) tissues.Expression of seven common endogenous control genes (glyceraldehyde-3-phosphate dehydrogenase (GAPDH), v-abl Abelson murine leukaemia viral oncogene homologue 1, beta-2-microglobulin, hypoxanthin phosphoribosyltransferase 1 (HPRT1), phosphoglycerate kinase 1, peptidylprolyl isomerase A, and ribosomal protein, large, P0) in 18 heterogenous NSCLC tumour specimens, 10 normal lung tissues and six NSCLC cell lines were analysed by quantitative RTPCR. The variances and correlation coefficients of cycle threshold (Ct) value of each control gene in three tissue groups and subgroups were compared. The difference and correlation coefficients between the Ct value for each control gene and the mean Ct value of the remaining control genes were calculated.The GAPDH gene transcript showed the least variance and linear regression analysis demonstrated that GAPDH and HPRT had the strongest correlation in pooled tumour and normal lung tissues. Furthermore, GAPDH expression value showed stringent correlation and had the lowest difference with the mean expression value of the remaining endogenous control genes.Among the seven common endogenous control genes, glyceraldehyde-3-phosphate dehydrogenase is the most suitable for quantitative RT-PCR reaction in nonsmall cell lung cancer tissue samples.