Normalization of Gene Expression for Acute Leukemia Studies: Selecting a Set of Endogenous Genes Control

Abstract

Reference genes are used as internal reaction control for gene expression analysis and for this they are considered reliable and must meet several important criteria. In view of the absence of studies regarding the best reference gene for the analysis of acute leukemia patients, a panel of genes commonly used as endogenous was selected from the literature for analysis of stability: GAPDH, ABL, HPRT1, RPLP0, ACTB and TBP. The stability of candidate reference genes was analyzed according to four statistical methods of assessment, namely, NormFinder, GeNorm and R software. From this study’s analysis it was possible to identify that the endogenous set composed by ACTB, ABL, TBP and RPLPO demonstrated good performances and stable expressions between the analyzed groups. Besides that, the GAPDH and HPRT genes could not be classified as good reference genes, considering that they presented a high standard deviation and great variability between groups, indicating low stability. Given these findings, this study suggests the main endogenous set for use as control/reference for the analysis of gene expression in peripheral blood and bone marrow samples from patients with acute leukemias, is composed by the ACTB, ABL, TBP and RPLPO genes.