大鼠管家基因的广泛的性别和/或激素依赖性表达。
Extensive sex- and/or hormone-dependent expression of rat housekeeping genes.
机构信息
Laboratories of Biochemistry, University of Pennsylvania, School of Veterinary Medicine, Philadelphia, PA 19104, USA.
出版信息
Endocr Res. 2013;38(2):105-11. doi: 10.3109/07435800.2012.723294. Epub 2012 Sep 19.
AIM
Identify sex- and hormone-independent housekeeping genes in rat liver by using a commercially available quantitative reverse transcription-polymerase chain reaction array designed to measure the expression of 32 rat housekeeping genes.
RESULTS
We found that the levels of five of the genes were sexually dimorphic, 22 genes were overexpressed, and one was underexpressed in multi-hormone-deficient hypophysectomized rats of both sexes. Only three genes fulfilled the stability criteria determined by geNorm and NormFinder as suitable housekeeping genes. Normalizing quantitative reverse transcription-polymerase chain reaction data with either of these three genes alone, the geometric means of any two of the genes, or even the geometric mean of all the three genes, produced similar results. In contrast, application of unproven housekeeping genes could lead to erroneous conclusions, having found that insulin-like growth factor 1 messenger RNA levels could be calculated dramatically either as male or as female predominant depending on the choice of housekeeping gene.
CONCLUSION
It is essential to validate the constancy of housekeeping genes under every experimental condition. (This research protocol was approved by the university's Institutional Animal Care and Use Committee.).
目的
通过使用商业定量逆转录-聚合酶链反应（qRT-PCR）芯片来测量 32 个大鼠管家基因的表达，确定大鼠肝脏中性别和激素无关的管家基因。
结果
我们发现，五个基因在性别上存在差异，22 个基因在两性去势多激素缺乏大鼠中表达过度，一个基因表达不足。只有三个基因符合 geNorm 和 NormFinder 确定的稳定性标准，适合作为管家基因。使用这三个基因中的任意两个或三个基因的几何平均值单独对 qRT-PCR 数据进行归一化，会产生相似的结果。相比之下，使用未经证实的管家基因可能会导致错误的结论，我们发现，胰岛素样生长因子 1 信使 RNA 水平的计算结果可能会根据管家基因的选择，显著地偏向男性或女性。
结论
在每个实验条件下，验证管家基因的稳定性是至关重要的。（本研究方案已获得大学机构动物护理和使用委员会的批准。）
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