Reference gene selection for head and neck squamous cell carcinoma gene expression studies.
Reference gene selection for head and neck squamous cell carcinoma gene expression studies.
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DOI：
10.1186/1471-2199-10-78
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发表时间：
2009-08-03
影响因子：
--
通讯作者：
Brouillet JP
中科院分区：
文献类型：
--
作者：
Lallemant B;Evrard A;Combescure C;Chapuis H;Chambon G;Raynal C;Reynaud C;Sabra O;Joubert D;Hollande F;Lallemant JG;Lumbroso S;Brouillet JP
盲目地选择内参基因已经不够了。我们首次提出了一项研究，确定了12种常用的癌症研究内参基因（ACT、ALAS、B2M、GAPDH、HMBS、HPRT、KALPHA、RPS18、RPL27、RPS29、SHAD和TBP）在头颈部鳞状细胞癌（HNSCC）领域定量表达数据规范化中的适用性。采用RT-qPCR方法检测46例HNSCC和正常匹配粘膜的原始表达水平。我们使用geNorm和NormFinder分析表达稳定性，并比较亚组之间的表达水平。在HNSCC和/或正常粘膜中，4个最佳的归一化基因是ALAS、GAPDH、RPS18和SHAD，两个基因最稳定的组合是GAPDH-SHAD。我们推荐在T1-T2肿瘤中使用KALPHA-TBP，在T3-T4肿瘤中使用RPL27-SHAD，在N0肿瘤中使用KALPHA-SHAD，在N+肿瘤中使用ALAS-TBP。ACT、B2M、GAPDH、HMBS、HPRT、KALPHA、RPS18、RPS29、SHAD和TBP在肿瘤和正常粘膜之间有轻微的失调（<1.7倍），但根据检测所需的分辨率，可以用于正常化。在HNSCC领域，本研究将指导研究人员根据实验的具体设置，从12个可能合适的内参基因中选择最合适的内参基因。
It is no longer adequate to choose reference genes blindly. We present the first study that defines the suitability of 12 reference genes commonly used in cancer studies (ACT, ALAS, B2M, GAPDH, HMBS, HPRT, KALPHA, RPS18, RPL27, RPS29, SHAD and TBP) for the normalization of quantitative expression data in the field of head and neck squamous cell carcinoma (HNSCC). Raw expression levels were measured by RT-qPCR in HNSCC and normal matched mucosa of 46 patients. We analyzed the expression stability using geNorm and NormFinder and compared the expression levels between subgroups. In HNSCC and/or normal mucosa, the four best normalization genes were ALAS, GAPDH, RPS18 and SHAD and the most stable combination of two genes was GAPDH-SHAD. We recommend using KALPHA-TBP for the study of T1-T2 tumors, RPL27-SHAD for T3-T4 tumors, KALPHA-SHAD for N0 tumors, and ALAS-TBP for N+ tumors. ACT, B2M, GAPDH, HMBS, HPRT, KALPHA, RPS18, RPS29, SHAD and TBP were slightly misregulated (<1.7-fold) between tumor and normal mucosa but can be used for normalization, depending on the resolution required for the assay. In the field of HNSCC, this study will guide researchers in selecting the most appropriate reference genes from among 12 potentially suitable reference genes, depending on the specific setting of their experiments.
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鉴定乳腺癌中实时RT-PCR数据归一化的基因。
影响因子：
3.8
作者：
Lyng MB;Laenkholm AV;Pallisgaard N;Ditzel HJ
通讯作者：
Ditzel HJ
影响因子：
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作者：
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4.1
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作者：
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通讯作者：
Füzesi, L
影响因子：
2.9
作者：
Johansson, Sofia;Fuchs, Andrea;Ekstrom, Tomas J.
通讯作者：
Ekstrom, Tomas J.