Selection of Novel Reference Genes for qPCR Normalisation in the MDA-MB-436 Breast Cancer Cell Line

Nowadays reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) is a widely used laboratory method for evaluating gene expression and its differences in various conditions. However, to increase the credibility and accuracy of the results, they are usually normalised against a pair or a triplet of reference genes in which expression is found to be stable across the conditions investigated in the study. This study aims to find suitable reference genes for evaluating gene expression in different confluences in the triple-negative breast cancer cell line MDAMB-436. A candidate reference gene set comprised of 30 genes was analysed in the study. The expression of these genes and four different genes of interest was analysed in over 90 different samples from the cell line MDA-MB-436 and then the most stable reference genes were determined using algorithms such as NormFinder, RefFinder, etc. All reference genes exhibited robust stability while the most stable reference genes were found to be MYL12A; RBX1; PUM1 and PFN1. The results of this study will provide other researchers around the world with knowledge about the most stable reference genes across different confluences in cell line MDA-MB-436 and will facilitate further research in this field.