Housekeeping genes as internal standards in cancer research.
Housekeeping genes as internal standards in cancer research.
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DOI：
10.2165/00066982-200408020-00005
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发表时间：
2004-01-01
期刊：
影响因子：
--
通讯作者：
Bakker, Annette
中科院分区：
文献类型：
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作者：
Janssens, Nico;Janicot, Michel;Bakker, Annette
背景：基因表达的差异在恶性组织中经常遇到，并且已经被深入研究，因为它们可以反映不同的实验或临床条件。通过与内源对照的表达进行比较，对信使 RNA 含量中通常微妙的变化进行量化。这些内源对照（例如看家基因）的适当选择对于有意义的定量 RNA 分析至关重要。看家基因最重要的特征是它们存在于所有细胞中，并且它们的表达水平在不同的实验条件下保持相对恒定。然而，没有一个管家基因在所有实验条件下总是表现出稳定的表达水平。因此，有必要在测试转录效应的特定实验条件下表征各种管家基因作为内部 RNA 对照的适用性。目的：本研究的目的是确定一些管家基因在癌症研究中用作内部标准的有效性。方法：将管家基因胆色素原脱氨酶 (PBGD) 和线粒体 ATP 合酶 6 (mATPsy6) 的表达与更常用的表达进行比较。 3-磷酸甘油醛脱氢酶（GAPDH）。我们使用实时定量 (RTq)-PCR 检查了许多细胞系和肿瘤与匹配的正常组织样本。结果：我们的研究结果表明，在细胞系中，所有三个所研究的管家基因都可以用作内部对照。当将肿瘤组织样本与匹配的正常组织样本进行比较时，我们验证了线粒体 ATPsy6 (mATPsy6) 是管家基因的最佳选择。结论：由于基因表达研究在临床环境中变得越来越重要，特别是在癌症诊断和治疗中，因此在这些研究中使用可靠的管家基因来标准化基因表达至关重要。我们的结论是，在解释涉及基因表达定量的实验时，管家基因的错误选择可能会导致错误。我们的研究证明了在比较肿瘤组织样本与正常组织样本时 mATPsy6 作为内源性对照的有用性。
BACKGROUND: Differences in gene expression are frequently encountered in malignant tissues, and have been intensively studied as they can reflect different experimental or clinical conditions. Quantification of the often subtle changes in messenger RNA content is performed through comparison with the expression of endogenous controls. The appropriate choice of these endogenous controls (e.g. housekeeping genes) is critical for meaningful quantitative RNA analysis. The most important characteristics of housekeeping genes are that they are present in all cells and that their expression levels remain relatively constant in different experimental conditions. However, no single housekeeping gene always manifests stable expression levels under all experimental conditions. Therefore, it is necessary to characterize the suitability of various housekeeping genes to serve as internal RNA controls under particular experimental conditions where transcription effects are being tested.AIM: It was the aim of this study to determine the validity of a number of housekeeping genes for their use as internal standards in cancer research.METHODS: The expression of the housekeeping genes porphobilinogen deaminase (PBGD) and mitochondrial ATP synthase 6 (mATPsy6), were compared with the expression of the more commonly used glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We examined a number of cell lines and tumor versus matched normal tissue samples using real-time quantitative (RTq)-PCR.RESULTS: Our findings suggest that in cell lines, all three of the studied housekeeping genes can be used as an internal control. When comparing tumor tissue samples with matched normal tissue samples, we validated mitochondrial ATPsy6 (mATPsy6) as the best choice for a housekeeping gene.CONCLUSION: Since gene expression studies are becoming increasingly important in the clinical environment, especially in cancer diagnosis and treatment, the use of an reliable housekeeping gene in these studies to normalize gene expression is essential. We conclude that a bad choice of housekeeping gene may lead to errors when interpreting experiments involving quantitation of gene expression. Our study demonstrated the usefulness of mATPsy6 as an endogenous control when comparing tumor tissue samples with normal tissue samples.