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Validation of Endogenous Control Genes by Real-Time Quantitative Reverse Transcriptase Polymerase Chain Reaction for Acute Leukemia Gene Expression Studies.
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.
Afiliação
  • Pessoa FMCP; Department of Medicine, Pharmacogenetics Laboratory, Drug Research and Development Center (NPDM), Federal University of Ceará, Fortaleza 60430-275, CE, Brazil.
  • Viana VBJ; Department of Biological Sciences, Oncology Research Center, Federal University of Pará, Belém 66073-005, PA, Brazil.
  • de Oliveira MB; Department of Biological Sciences, Oncology Research Center, Federal University of Pará, Belém 66073-005, PA, Brazil.
  • Nogueira BMD; Department of Medicine, Pharmacogenetics Laboratory, Drug Research and Development Center (NPDM), Federal University of Ceará, Fortaleza 60430-275, CE, Brazil.
  • Ribeiro RM; Department of Hematology, Fortaleza General Hospital (HGF), Fortaleza 60150-160, CE, Brazil.
  • Oliveira DS; Department of Medicine, Pharmacogenetics Laboratory, Drug Research and Development Center (NPDM), Federal University of Ceará, Fortaleza 60430-275, CE, Brazil.
  • Lopes GS; Department of Hematology, Fortaleza General Hospital (HGF), Fortaleza 60150-160, CE, Brazil.
  • Vieira RPG; Department of Hematology, César Cals General Hospital, Fortaleza 60015-152, CE, Brazil.
  • de Moraes Filho MO; Department of Hematology, São Vicente de Paulo Maternity Hospital, Barbalha 63180-000, CE, Brazil.
  • de Moraes MEA; Department of Medicine, Pharmacogenetics Laboratory, Drug Research and Development Center (NPDM), Federal University of Ceará, Fortaleza 60430-275, CE, Brazil.
  • Khayat AS; Department of Medicine, Pharmacogenetics Laboratory, Drug Research and Development Center (NPDM), Federal University of Ceará, Fortaleza 60430-275, CE, Brazil.
  • Moreira FC; Department of Biological Sciences, Oncology Research Center, Federal University of Pará, Belém 66073-005, PA, Brazil.
  • Moreira-Nunes CA; Department of Biological Sciences, Oncology Research Center, Federal University of Pará, Belém 66073-005, PA, Brazil.
Genes (Basel) ; 15(2)2024 01 24.
Article em En | MEDLINE | ID: mdl-38397141
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), ß-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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Leucemia / Perfilação da Expressão Gênica Limite: Animals / Humans Idioma: En Revista: Genes (Basel) Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Suíça
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Leucemia / Perfilação da Expressão Gênica Limite: Animals / Humans Idioma: En Revista: Genes (Basel) Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Suíça