Different stability of miRNAs and endogenous control genes in archival specimens of papillary thyroid carcinoma

Molecular Medicine

Valdas Sarauskas¹  Albertas Dauksa²  Daina Pamedytyte³  Aurelija Zvirbliene³  Enrika Leipute³  Birute Zilaitiene⁴  Dalia Dauksiene⁴ 

[1] Department of Pathology, Lithuanian University of Health Sciences, Eiveniu Str. 2, 50161, Kaunas, Lithuania;Institute for Digestive Research, Medical Academy, Lithuanian University of Health Sciences, Eiveniu Str. 2, 50161, Kaunas, Lithuania;Institute of Biotechnology, Life Sciences Center, Vilnius University, Sauletekio Av. 7, 10257, Vilnius, Lithuania;Institute of Endocrinology, Medical Academy, Lithuanian University of Health Sciences, Eiveniu Str. 2, 50161, Kaunas, Lithuania;

关键词: Stability;    miRNA;    Endogenous control;    FFPE;    Papillary thyroid carcinoma;   

DOI  :  10.1186/s10020-020-00218-7

来源: Springer

【 摘 要 】

BackgroundThe most popular miRNA quantitation technique is RQ-PCR with relative gene expression method that requires an endogenous control (EC) gene for data normalization. However, there are insufficient data and selection criteria on the most suitable ECs for miRNA expression studies in many cancer types including papillary thyroid carcinoma (PTC). Therefore, in this study we evaluated the impact of chosen EC and archival formalin-fixed, paraffin-embedded (FFPE) PTC tissue age on estimated target miRNA expression.MethodsRQ-PCR was used to determine expression levels of five miRNAs (miR-146b, miR-222, miR-21, miR-221 and miR-181b) and three different endogenous controls (RNU48, let-7a, miR-16), which were used to normalize the data. In total, 400 FFPE PTC tissues were analyzed that have been stored from 1 to 15 years.ResultsThe stability of commonly used ECs RNU48 and let-7a significantly differs from the stability of target miRNA in archival FFPE PTC tissues. Moreover, these differences have a great impact on miRNA expression results when FFPE tissue samples have been stored for a different period of time.ConclusionsIt is important to select an ECs not only stable in the tissue of interest but also with similar stability to target miRNA, especially when working with samples of different age.

【 授权许可】

CC BY