Identification of reference genes and miRNAs for RT-qPCR in testosterone propionate-induced benign prostatic hyperplasia in rats.

Abstract

Testosterone propionate-induced benign prostatic hyperplasia in rats is a common model that is widely used in studies of the effects and molecular mechanisms of drugs designed to treat benign prostatic hyperplasia. RT-qPCR is a widely used technique in gene expression studies. Proper normalisation is critical for accurate expression analysis. Currently, no validated reference genes are available for RT-qPCR in rat benign prostatic hyperplasia. Given that microRNAs regulate mRNA expression at the post-transcriptional level, they are usually studied together. Here, the expression stability of 21 putative reference genes including 8 mRNAs and 13 miRNAs was evaluated in benign prostatic hyperplasia in rats. Relative expression levels of each gene were detected in rats from a model group and a normal group using SYBR RT-qPCR. Expression stability was evaluated by geNorm and NormFinder. The commonly used reference genes, such as ACTB, B2M and mir-16, were less stable, and let-7a was eliminated due to a large Ct value, most likely indicating a relatively low expression level. Therefore, to obtain reliable results, mir-26a was recommended as a suitable reference for miRNA expression analysis and EF-1a as a suitable reference for mRNA analysis in testosterone propionate-induced benign prostatic hyperplasia in rats.