Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury.
Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury.
复制标题
选择适合脊髓损伤大鼠基因表达正常化的内部对照
DOI：
10.4103/1673-5374.327350
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发表时间：
2022-06
影响因子：
6.1
通讯作者：
Wang YX
中科院分区：
文献类型：
--
作者：
Liu W;Yu J;Wang YF;Shan QQ;Wang YX
在脊髓损伤研究中，实时荧光逆转录聚合酶链反应用于基因表达正常化的内参基因表达缺乏系统研究。在这项研究中，我们使用大鼠脊髓半横断模型来分析13个常用参考基因的表达稳定性：Actb，Ankrd 27，CypA，Gapdh，Hprt 1，Mrpl 10，Pgk 1，Rictor，Rn 18 s，Tbp，Ubc，Ubxn 11和Ywhaz。我们的结果表明，Ankrd 27，Ubc和Tbp的表达是稳定的脊髓损伤后，而Actb是最不稳定的内部控制基因。因此，使用Ankrd 27、Ubc、Tbp和Actb来研究具有不同稳定性的内部对照基因对靶基因表达的正常化的影响。当Ankrd 27、Ubc和Tbp用作内部对照时，靶基因表达水平和随时间的变化相似，但当Actb用作内部对照时不同。我们建议Ankrd 27，Ubc和Tbp被用作实时逆转录聚合酶链反应在脊髓损伤研究的内部控制基因。本研究于2018年3月4日获得中国江苏省实验动物管理委员会批准（批准号：20180304-008）。
There is a lack of systematic research on the expression of internal control genes used for gene expression normalization in real-time reverse transcription polymerase chain reaction in spinal cord injury research. In this study, we used rat models of spinal cord hemisection to analyze the expression stability of 13 commonly applied reference genes: Actb, Ankrd27, CypA, Gapdh, Hprt1, Mrpl10, Pgk1, Rictor, Rn18s, Tbp, Ubc, Ubxn11, and Ywhaz. Our results show that the expression of Ankrd27, Ubc, and Tbp were stable after spinal cord injury, while Actb was the most unstable internal control gene. Ankrd27, Ubc, Tbp, and Actb were consequently used to investigate the effects of internal control genes with differing stabilities on the normalization of target gene expression. Target gene expression levels and changes over time were similar when Ankrd27, Ubc, and Tbp were used as internal controls but different when Actb was used as an internal control. We recommend that Ankrd27, Ubc, and Tbp are used as internal control genes for real-time reverse transcription polymerase chain reaction in spinal cord injury research. This study was approved by the Administration Committee of Experimental Animals, Jiangsu Province, China (approval No. 20180304-008) on March 4, 2018.
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