人脊索瘤中内参基因的验证
Validation of reference genes in human chordoma.
作者信息
Santegoeds R G C, Yakkioui Y, Jahanshahi A, Hoogland G, Temel Y, van Overbeeke J J
机构信息
Department of Neurosurgery, School for Mental Health and Neuroscience, Maastricht University Medical Centre, Maastricht, The Netherlands.
出版信息
Surg Neurol Int. 2017 Jun 5;8:100. doi: 10.4103/sni.sni_399_16. eCollection 2017.
BACKGROUND
Chordoma are rare slow-growing tumors of the axial skeleton, which are thought to arise from remnants of the notochord. Little is known about the underlying mechanisms that drive this tumor. However, the assessment of gene expression levels by quantitative real-time polymerase chain reaction (qRT-PCR) is hampered due to a lack of validated reference genes. Using an unstable reference gene in qRT-PCR may lead to irreproducible results.
METHODS
The expression of 12 candidate reference genes (, , , , , , , , , , , and ) was analyzed by qRT-PCR in flash frozen chordoma samples from 18 patients. GeNorm and NormFinder algorithms were used to rank the stability of the genes.
RESULTS
From most to least stably expressed, the top six genes found by geNorm were , , , , , and . When analyzed by NormFinder, the top six genes were , , , , , and . alone, which is often used as a reference gene in chordoma gene expression studies, is not stable enough for reliable results.
CONCLUSION
In gene expression studies of human chordomas, , , and are more stably expressed, and therefore, are preferred reference genes over the most often used reference gene so far, .
背景
脊索瘤是罕见的生长缓慢的中轴骨肿瘤，被认为起源于脊索的残余部分。对于驱动这种肿瘤的潜在机制知之甚少。然而，由于缺乏经过验证的内参基因，通过定量实时聚合酶链反应（qRT-PCR）评估基因表达水平受到阻碍。在qRT-PCR中使用不稳定的内参基因可能会导致结果不可重复。
方法
通过qRT-PCR分析了18例患者的冷冻脊索瘤样本中12个候选内参基因（、、、、、、、、、、和）的表达。使用GeNorm和NormFinder算法对基因的稳定性进行排名。
结果
GeNorm发现的最稳定到最不稳定表达的前六个基因是、、、、、和。通过NormFinder分析时，前六个基因是、、、、、和。单独来看，在脊索瘤基因表达研究中经常用作内参基因的它，稳定性不足以获得可靠结果。
结论
在人类脊索瘤的基因表达研究中，、和表达更稳定，因此，相对于迄今为止最常用的内参基因，它们是更优的内参基因。
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