Validation of reference genes for normalizing gene expression in real-time quantitative reverse transcription PCR in human thyroid cells in primary culture treated with progesterone and estradiol.
The use of appropriately chosen reference genes for normalizing gene expression in real-time quantitative reverse transcription polymerase chain reaction is an important step in the analysis of gene expression, compensating for several technical factors. As female sex hormones have been shown to influence growth and differentiation of thyroid follicular cells, the establishment of normalizer genes in human thyroid cells in primary culture, treated with progesterone, and estradiol, is important to evaluate their effect on gene expression in these cells, so candidate reference genes were studied. β-Actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β2-microglobulin (B2M), and TATA box binding protein (TBP) were evaluated in thyroid cells treated with estradiol, progesterone, and their inhibitors. Normfinder software was used to assess the stability of the genes and identified β-actin as the gene with adequate stability and lower inter-group variations, when compared to TBP, B2M, and GAPDH.
Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, Porto Alegre, Brazil.
Ana Paula
AP
Santin
Aline Francielle Damo
AF
Souza
Llma Simoni
LS
Brum
Tania Weber
TW
Furlanetto
D004958:Estradiol / Q000494:pharmacology*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D011374:Progesterone / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism*
D011374:Progesterone; D004958:Estradiol