Validation of Reference Genes for Normalization Gene Expression in Reverse Transcription Quantitative PCR in Human Normal Thyroid and Goiter Tissue

Abstract EN

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.