Validation of reference genes in human epicardial adipose tissue and left ventricular myocardium in heart failure

Abstract

The characterization of tissue-specific gene expression in failing hearts requires the identification of appropriately validated reference genes. This study sought to validate which commonly used reference genes were the most suitable for qRT-PCR data normalization in epicardial adipose tissue and left ventricular myocardium in patients with end-stage heart failure. The mRNA expression of 10 candidate reference genes was analyzed by qRT-PCR. The stability of their expression was evaluated by our novel scoring system and compared with known algorithms used to identify stable reference genes (Normfinder and Bestkeeper). Using these three methods, we selected the Ribosomal protein L13A (RPL13A) as the best reference gene when studying either epicardial adipose tissue or left ventricular myocardium in failing hearts. In contrast, the β2-microglobulin (B2M) gene was identified as a more suitable reference gene in studies comparing gene expression in epicardial adipose tissue to left ventricular myocardium.