Identification of genes for normalization of real-time RT-PCR data in breast carcinomas.
Identification of genes for normalization of real-time RT-PCR data in breast carcinomas.
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鉴定乳腺癌中实时RT-PCR数据归一化的基因。
DOI：
10.1186/1471-2407-8-20
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发表时间：
2008-01-22
期刊：
影响因子：
3.8
通讯作者：
Ditzel HJ
中科院分区：
文献类型：
--
作者：
Lyng MB;Laenkholm AV;Pallisgaard N;Ditzel HJ
实时定量RT-PCR（RT-qPCR）已成为基础和转化生物医学研究中一种有价值的分子技术，并正在成为一种同样有价值的临床工具。样本间值的相关性需要数据归一化，这可以通过各种手段来实现，其中最常见的是对内部稳定表达的参考基因的归一化。最近，已经发现传统上使用的参考基因如GAPDH和B2 M在不同组织中的各种情况下受到调节，强调了鉴定与影响组织的因素无关的基因并且在实验环境中稳定表达的基因的需要。在这项研究中，我们鉴定了用于浸润性乳腺癌（IBC）的RT-qPCR数据标准化的基因，特别强调雌激素受体阳性（ER+）IBC，但也检查了它们对ER-IBC、正常乳腺组织和乳腺癌细胞系的适用性。通过qRT-PCR研究的参考基因是RPLP 0、TBP、ABL 1、ACTB、GUS-B、ABL 1、GAPDH和B2 M。检查了18个切除的组织标本（11个ER+ IBC，4个ER-IBC，3个正常乳腺组织）和3个ER+细胞系的活检，并通过描述性统计、geNorm和Normalt分析数据。此外，选择的参考基因在激光捕获显微切割ER+ IBC细胞的表达进行了比较，与整个组织。通过GeNorm对人组织样品的组合组（ER+和ER-IBC以及正常乳腺组织）和浸润性癌样品（ER+和ER-IBC）两者鉴定了一组3个基因TBP、RPLP 0和RPLP 1，其中Norm一致地鉴定了所有样品组合的单一最佳基因RPLP 1。当对正常和恶性乳腺标本进行RT-qPCR时，选择的参考基因应该是收集的3个基因（TBP、RPLP 0和RPLP 1）的平均值，或RPLP 1作为单个基因。
Quantitative real-time RT-PCR (RT-qPCR) has become a valuable molecular technique in basic and translational biomedical research, and is emerging as an equally valuable clinical tool. Correlation of inter-sample values requires data normalization, which can be accomplished by various means, the most common of which is normalization to internal, stably expressed, reference genes. Recently, such traditionally utilized reference genes as GAPDH and B2M have been found to be regulated in various circumstances in different tissues, emphasizing the need to identify genes independent of factors influencing the tissue, and that are stably expressed within the experimental milieu. In this study, we identified genes for normalization of RT-qPCR data for invasive breast cancer (IBC), with special emphasis on estrogen receptor positive (ER+) IBC, but also examined their applicability to ER- IBC, normal breast tissue and breast cancer cell lines. The reference genes investigated by qRT-PCR were RPLP0, TBP, PUM1, ACTB, GUS-B, ABL1, GAPDH and B2M. Biopsies of 18 surgically-excised tissue specimens (11 ER+ IBCs, 4 ER- IBCs, 3 normal breast tissues) and 3 ER+ cell lines were examined and the data analyzed by descriptive statistics, geNorm and NormFinder. In addition, the expression of selected reference genes in laser capture microdissected ER+ IBC cells were compared with that of whole-tissue. A group of 3 genes, TBP, RPLP0 and PUM1, were identified for both the combined group of human tissue samples (ER+ and ER- IBC and normal breast tissue) and for the invasive cancer samples (ER+ and ER- IBC) by GeNorm, where NormFinder consistently identified PUM1 at the single best gene for all sample combinations. The reference genes of choice when performing RT-qPCR on normal and malignant breast specimens should be either the collected group of 3 genes (TBP, RPLP0 and PUM1) employed as an average, or PUM1 as a single gene.
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影响因子：
2.9
作者：
Haller, F;Kulle, B;Füzesi, L
通讯作者：
Füzesi, L
影响因子：
2.9
作者：
Morse, DL;Carroll, D;Gillies, RJ
通讯作者：
Gillies, RJ
影响因子：
2.9
作者：
Johansson, Sofia;Fuchs, Andrea;Ekstrom, Tomas J.
通讯作者：
Ekstrom, Tomas J.
DOI：
10.1016/j.jsbmb.2005.04.022
发表时间：
2005-05-01
影响因子：
4.1
作者：
Dowsett, M;Martin, LA;Johnston, S
通讯作者：
Johnston, S
影响因子：
5
作者：
Antonov, J;Goldstein, DR;Jaggi, R
通讯作者：
Jaggi, R