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- Title
Validation of Endogenous Control Genes by Real-Time Quantitative Reverse Transcriptase Polymerase Chain Reaction for Acute Leukemia Gene Expression Studies.
- Authors
Pessoa, Flávia Melo Cunha de Pinho; Viana, Vitória Beatriz de Jesus; de Oliveira, Marcelo Braga; Nogueira, Beatriz Maria Dias; Ribeiro, Rodrigo Monteiro; Oliveira, Deivide de Sousa; Lopes, Germison Silva; Vieira, Ricardo Parente Garcia; de Moraes Filho, Manoel Odorico; de Moraes, Maria Elisabete Amaral; Khayat, André Salim; Moreira, Fabiano Cordeiro; Moreira-Nunes, Caroline Aquino
- Abstract
Reference genes are used as internal reaction controls for gene expression analysis, and for this reason, 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 controls was selected from the literature for stability analysis: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Abelson murine leukemia viral oncogene human homolog 1 (ABL), Hypoxanthine phosphoribosyl-transferase 1 (HPRT1), Ribosomal protein lateral stalk subunit P0 (RPLP0), β-actin (ACTB) and TATA box binding protein (TBP). The stability of candidate reference genes was analyzed according to three statistical methods of assessment, namely, NormFinder, GeNorm and R software (version 4.0.3). From this study's analysis, it was possible to identify that the endogenous set composed of ACTB, ABL, TBP and RPLP0 demonstrated good performances and stable expressions between the analyzed groups. In addition to 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 gene set for use as a control/reference for the gene expression in peripheral blood and bone marrow samples from patients with acute leukemias is composed of the ACTB, ABL, TBP and RPLP0 genes. Researchers may choose two to three of these housekeeping genes to perform data normalization.
- Subjects
REVERSE transcriptase polymerase chain reaction; GENE expression; ACUTE leukemia; REAL-time control; MOUSE leukemia
- Publication
Genes, 2024, Vol 15, Issue 2, p151
- ISSN
2073-4425
- Publication type
Article
- DOI
10.3390/genes15020151