选择合适的内参基因以研究产前糖皮质激素给药对人胎盘影响的重要性。
The importance of selecting the right internal control gene to study the effects of antenatal glucocorticoid administration in human placenta.
作者信息
Gütling H, Bionaz M, Sloboda D M, Ehrlich L, Braun F, Gramzow A K, Henrich W, Plagemann A, Braun T
机构信息
Departments of Obstetrics and Division of Experimental Obstetrics, Charité, Berlin, Germany.
Animal and Rangeland Sciences, Oregon State University, Corvallis, USA.
出版信息
Placenta. 2016 Aug;44:19-22. doi: 10.1016/j.placenta.2016.05.011. Epub 2016 May 27.
RT-qPCR requires a suitable set of internal control genes (ICGs) for an accurate normalization. The usefulness of 7 previously published ICGs in the human placenta was analyzed according to the effects of betamethasone treatment, sex and fetal age. Raw RT-qPCR data of the ICGs were evaluated using published algorithms. The algorithms revealed that a reliable normalization was achieved using the geometrical mean of PPIA, RPL19, HMBS and SDHA. The use of a different subset ICGs out of the 7 investigated, although not statistically affected by the conditions, biased the results, as demonstrated through changes in expression of glucocorticoid receptor (NR3C1) mRNA as a target gene.
逆转录定量聚合酶链反应（RT-qPCR）需要一组合适的内参基因（ICGs）来进行准确的标准化。根据倍他米松治疗、性别和胎龄的影响，分析了7个先前发表的人胎盘内参基因的实用性。使用已发表的算法评估内参基因的原始RT-qPCR数据。这些算法表明，使用PPIA、RPL19、HMBS和SDHA的几何平均值可实现可靠的标准化。在所研究的7个内参基因中使用不同的子集，尽管不受这些条件的统计学影响，但会使结果产生偏差，这通过作为靶基因的糖皮质激素受体（NR3C1）mRNA表达的变化得到了证明。
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