人白细胞中腺苷受体表达及基因参考评估
Adenosine receptor expression and gene reference evaluation in human leukocytes.
作者信息
Sabatino Laura, Cabiati Manuela, Caselli Chiara, Del Ry Silvia
机构信息
CNR Institute of Clinical Physiology, Laboratory of Cardiovascular Biochemistry, Pisa, Italy.
出版信息
Clin Lab. 2013;59(5-6):571-7. doi: 10.7754/clin.lab.2012.120219.
BACKGROUND
Peripheral blood mononuclear cells and isolated polymorphonuclear neutrophilis were used to evaluate gene expression studies. Unfortunately, there are many methodological problems related to these cellular models, limiting their use. The aim was to evaluate a fast and easy procedure for the extraction of total RNA from leukocytes obtained from human whole blood (WB) < 10 mL; to determine adenosine receptor (AR) mRNA expression in WB samples of normal subjects and to establish the most stable reference genes for data normalization.
METHODS
mRNA expression was performed by Real-Time PCR.
RESULTS
The most stably expressed genes were TPT1, EEF1A, and RPL13A. Similar levels of mRNA expression were observed for A2aR, A2bR, and A3R while lower levels were measured for A1R (p = 0.02 A1R vs. A2aR; p = 0.04 A1R vs. A3R).
CONCLUSIONS
Our study represents an important and useful starting point for future investigations devoted to evaluate the expression of ARs in human diseases.
背景
外周血单个核细胞和分离的多形核中性粒细胞用于评估基因表达研究。不幸的是，这些细胞模型存在许多方法学问题，限制了它们的应用。目的是评估一种从小于10 mL人全血（WB）中获得的白细胞中快速简便提取总RNA的方法；确定正常受试者WB样本中腺苷受体（AR）mRNA的表达，并建立用于数据标准化的最稳定参考基因。
方法
通过实时荧光定量PCR进行mRNA表达检测。
结果
表达最稳定的基因是TPT1、EEF1A和RPL13A。A2aR、A2bR和A3R的mRNA表达水平相似，而A1R的表达水平较低（A1R与A2aR相比，p = 0.02；A1R与A3R相比，p = 0.04）。
结论
我们的研究为未来致力于评估人类疾病中ARs表达的研究提供了一个重要且有用的起点。
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