Gapdh Gene Expression Is Modulated by Inflammatory Arthritis and Is not Suitable for qPCR Normalization
Gapdh Gene Expression Is Modulated by Inflammatory Arthritis and Is not Suitable for qPCR Normalization
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DOI：
10.1007/s10753-014-9829-x
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发表时间：
2014-08-01
期刊：
影响因子：
5.1
通讯作者：
Perretti, Mauro
中科院分区：
文献类型：
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作者：
Montero-Melendez, Trinidad;Perretti, Mauro
基因表达研究是理解复杂疾病的基础，为诊断和患者分层提供了对致病过程的新见解和新工具。通过实时PCR进行的基因谱分析研究需要使用参考基因进行标准化，并且适当的验证对于准确的结果至关重要。我们对K/BxN血清诱导的小鼠关节炎模型中的六个常见管家基因进行了全面评估。经典统计学和Normandy分析指出Gapdh不太稳定，因此不适合作为参考对照。关节炎关节中Gapdh的表达显著下调，因此产生了对转录变化的高估。Hptr、B2 m和Rpll 3a显示最恒定的表达。总的来说，我们的数据建议在K/BxN模型的急性期的基因表达研究中不要使用Gapdh，并增加了一个警示说明，需要验证参考基因，以获得可靠、可比和可重复的结果。
Gene expression studies are fundamental for the understanding of complex diseases, providing new insights into the pathogenic process and new tools for diagnostic and patient stratification. Gene profiling studies by real-time PCR require the use of reference genes for normalization and an appropriate validation is essential for accurate results. We performed a comprehensive assessment of six common housekeeping genes in the K/BxN serum-induced arthritis model in mice. Classical statistics and NormFinder analyses pointed out Gapdh as the less stable and therefore unsuitable as a reference control. Gapdh was considerably down-regulated in arthritic joints and therefore produced an overestimation of transcriptional changes. Hptr, B2m, and Rpl13a showed the most constant expression. Collectively our data advise against the use of Gapdh in gene expression studies in the acute phase of the K/BxN model and adds a cautionary note on the need to validate the reference genes for reliable, comparable, and reproducible results.