Endogenous controls of gene expression in N-methyl-N-nitrosourea-induced T-cell lymphoma in p53-deficient mice.
Endogenous controls of gene expression in N-methyl-N-nitrosourea-induced T-cell lymphoma in p53-deficient mice.
复制标题
p53缺陷小鼠中N-甲基-N-亚硝基脲诱导的T细胞淋巴瘤基因表达的内源控制
DOI：
10.1186/s12885-017-3536-6
复制
发表时间：
2017-08-14
期刊：
影响因子：
3.8
通讯作者：
Fan C
中科院分区：
文献类型：
--
作者：
Wu X;Liu S;Lyu J;Zhou S;Yang Y;Wang C;Gu W;Zuo Q;Li B;Fan C
背景实时聚合酶链式反应 (PCR) 已成为基因表达谱分析中越来越重要的技术，因为它可以提供对复杂生物和病理过程的深入了解，并可用于预测疾病或治疗结果。尽管归一化数据对于准确估计 mRNA 表达水平是必要的，但一些证据表明所谓的看家基因的表达并不稳定。本研究旨在验证 N-甲基-N-亚硝基脲 (MNU) 诱导的 T 细胞淋巴瘤小鼠模型中实时 PCR 标准化的参考基因。方法通过用 37.5 mg/kg MNU 治疗，在 p53 缺陷小鼠中产生 T 细胞淋巴瘤。胸腺和脾脏被确定为淋巴瘤发病率最高的主要靶器官。我们分析了八个潜在内源参考基因（Gapdh、Rn18s、Actb、Hprt、B2M、Rplp0、Gusb、Ctbp1）的 RNA 表达水平。使用geNorm和NormFinder算法测试MNU治疗后不同时间点这些参考基因的表达稳定性。结果总共65%的MNU治疗小鼠出现T细胞淋巴瘤，其中脾脏和胸腺是主要靶器官。通过定量逆转录聚合酶链反应（RT-qPCR）有效扩增所有候选参考基因。 MNU治疗后和淋巴瘤发生过程中的基因稳定性评估显示，Ctbp1和Rplp0分别是胸腺和脾脏中表达最稳定的基因。使用另外两组引物对胸腺 RNA 进行 RT-PCR，证实 Ctbp1 是所有候选参考基因中最稳定的。结论我们为 T 细胞淋巴瘤模型中的基因表达研究提供了合适的内源对照。
BackgroundReal-time polymerase chain reaction (PCR) has become an increasingly important technique for gene expression profiling because it can provide insights into complex biological and pathological processes and be used to predict disease or treatment outcomes. Although normalized data are necessary for an accurate estimation of mRNA expression levels, several pieces of evidence suggest that the expression of so-called housekeeping genes is not stable. This study aimed to validate reference genes for the normalization of real-time PCR in an N-methyl-N-nitrosourea (MNU)-induced T-cell lymphoma mouse model.MethodsT-cell lymphomas were generated in p53-deficient mice by treatment with 37.5 mg/kg MNU. Thymus and spleen were identified as the primary target organs with the highest incidences of lymphomas. We analyzed the RNA expression levels of eight potential endogenous reference genes (Gapdh,Rn18s,Actb, Hprt,B2M,Rplp0,Gusb,Ctbp1). The expression stabilities of these reference genes were tested at different time points after MNU treatment using geNorm and NormFinder algorithms.ResultsA total of 65% of MNU-treated mice developed T-cell lymphomas, with the spleen and thymus as the major target organs. All candidate reference genes were amplified efficiently by quantitative reverse-transcription polymerase chain reaction (RT-qPCR). Gene stability evaluation after MNU treatment and during lymphomagenesis revealed thatCtbp1andRplp0were the most stably expressed genes in the thymus and spleen, respectively. RT-PCR of thymus RNA using two additional sets of primer confirmed thatCtbp1was the most stable of all the candidate reference genes.ConclusionsWe provided suitable endogenous controls for gene expression studies in the T-cell lymphoma model.
登录
查看更多内容
影响因子：
3.7
作者：
de Jonge HJ;Fehrmann RS;de Bont ES;Hofstra RM;Gerbens F;Kamps WA;de Vries EG;van der Zee AG;te Meerman GJ;ter Elst A
通讯作者：
ter Elst A
DOI：
10.2165/00066982-200408020-00005
发表时间：
2004-01-01
期刊：
Molecular diagnosis : a journal devoted to the understanding of human disease through the clinical application of molecular biology
影响因子：
--
作者：
Janssens, Nico;Janicot, Michel;Bakker, Annette
通讯作者：
Bakker, Annette
影响因子：
1.5
作者：
Takaoka, M;Sehata, S;Usui, T
通讯作者：
Usui, T
影响因子：
2.1
作者：
Meijerink, Jules P. P.
通讯作者：
Meijerink, Jules P. P.
DOI：
10.1186/1756-9966-29-144
发表时间：
2010-11-08
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
--
作者：
Sørby LA;Andersen SN;Bukholm IR;Jacobsen MB
通讯作者：
Jacobsen MB