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Adenosine Receptor Expression and Gene Reference Evaluation in Human Leukocytes by Laura Sabatino, Manuela Cabiati, Chiara Caselli, Silvia Del Ry

Background: Peripheral blood mononuclear cells and isolated polymorphonuclear neutrophilis were used to eval-uate 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) < 10mL; 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.

DOI: 10.7754/Clin.Lab.2012.120219