title: Validation of reference genes for normalization gene expression in reverse transcription quantitative PCR in human normal thyroid and goiter tissue.
abstract: Reverse transcription quantitative polymerase chain reaction (RT-qPCR) has been recognized as the most accurate method for quantifying mRNA transcripts, but normalization of samples is a prerequisite for correct data interpretation. So, this study aimed to evaluate the most stable reference gene for RT-qPCR in human normal thyroid and goiter tissues. Beta-actin (ACTB); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); succinate dehydrogenase, subunit A, flavoprotein (Fp) (SDHA); hypoxanthine phosphoribosyltransferase I (HPRTI); tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide (YWHAZ); and beta-2-microglobulin (B2M) were evaluated in 14 thyroid tissue samples (7 normal and 7 goiter tissues) by RT-qPCR. The mean Cq and the maximum fold change (MFC) and NormFinder software were used to assess the stability of the genes. As a result, ACTB gene was more stable than GAPDH, SDHA, HPRTI, YWHAZ, and B2M. In conclusion, ACTB could be used to normalize RT-qPCR data in normal thyroid and goiter tissues.
authors.affiliation: Programa de Pós Graduação em Medicina: Ciências Médicas, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos 2400, 90035-903 Porto Alegre, RS, Brazil.
authors.forename: Raquel
authors.identifier: 0000-0001-5083-9988
authors.initials: R
authors.lastname: Weber
authors.affiliation: Programa de Pós Graduação em Medicina: Ciências Médicas, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos 2400, 90035-903 Porto Alegre, RS, Brazil ; Programa de Pós Graduação em Ciências da Saúde, Universidade Federal de Ciências da Saúde Porto Alegre, Rua Sarmento Leite 245, 90050-170 Porto Alegre, RS, Brazil.
authors.forename: Ana Paula Santin
authors.initials: AP
authors.lastname: Bertoni
authors.affiliation: Departamento de Fisiologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite 500, 90050-170 Porto Alegre, RS, Brazil.
authors.forename: Laura Walter
authors.initials: LW
authors.lastname: Bessestil
authors.affiliation: Serviço de Patologia, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, 90035-903 Porto Alegre, RS, Brazil.
authors.forename: Beatriz Maria de Azevedo Assis
authors.initials: BM
authors.lastname: Brasil
authors.affiliation: Departamento de Fisiologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Sarmento Leite 500, 90050-170 Porto Alegre, RS, Brazil.
authors.forename: Llma Simoni
authors.initials: LS
authors.lastname: Brum
authors.affiliation: Programa de Pós Graduação em Medicina: Ciências Médicas, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos 2400, 90035-903 Porto Alegre, RS, Brazil.
authors.forename: Tania Weber
authors.initials: TW
authors.lastname: Furlanetto
mesh_terms: D015870:Gene Expression / Q000235:genetics*; D006042:Goiter / Q000235:genetics*; D006801:Humans; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013961:Thyroid Gland / Q000378:metabolism*