miR-16-5p Is a Stably-Expressed Housekeeping MicroRNA in Breast Cancer Tissues from Primary Tumors and from Metastatic Sites

International Journal of Molecular Sciences

Cornelia Hauser-Kronberger¹  Franz Zehentmayr²  Gerd Fastner²  Felix Sedlmayer²  Hubert Hackl³  Frank Hamacher⁴  Brigitte Mlineritsch⁴  Simon Peter Gampenrieder⁴  Richard Greil⁴  Clemens Hufnagl⁴  Gabriel Rinnerthaler⁴ 

[1] Department of Pathology, Paracelsus Medical University Salzburg, Salzburg 5020, Austria;Department of Radiotherapy, Paracelsus Medical University Salzburg, Salzburg 5020, Austria;Division of Bioinformatics, Biocenter, Medical University of Innsbruck, Innsbruck 6020, Austria;IIIrd Medical Department with Hematology and Medical Oncology, Oncologic Center, Paracelsus Medical University Salzburg, Salzburg 5020, Austria;

关键词: miR-16;    miR-16-5p;    microRNA;    endogenous control;    housekeeper;    control;    breast cancer;   

DOI  :  10.3390/ijms17020156

来源: DOAJ

【 摘 要 】

For quantitative microRNA analyses in formalin-fixed paraffin-embedded (FFPE) tissue, expression levels have to be normalized to endogenous controls. To investigate the most stably-expressed microRNAs in breast cancer and its surrounding tissue, we used tumor samples from primary tumors and from metastatic sites. MiRNA profiling using TaqMan® Array Human MicroRNA Cards, enabling quantification of 754 unique human miRNAs, was performed in FFPE specimens from 58 patients with metastatic breast cancer. Forty-two (72%) samples were collected from primary tumors and 16 (28%) from metastases. In a cross-platform analysis of a validation cohort of 32 FFPE samples from patients with early breast cancer genome-wide microRNA expression analysis using SurePrintG3 miRNA (8 × 60 K)® microarrays from Agilent® was performed. Eleven microRNAs could be detected in all samples analyzed. Based on NormFinder and geNorm stability values and the high correlation (rho ≥ 0.8) with the median of all measured microRNAs, miR-16-5p, miR-29a-3p, miR-126-3p, and miR-222-3p are suitable single gene housekeeper candidates. In the cross-platform validation, 29 human microRNAs were strongly expressed (mean log2-intensity > 10) and 21 of these microRNAs including miR-16-5p and miR-29a-3p were also stably expressed (CV < 5%). Thus, miR-16-5p and miR-29a-3p are both strong housekeeper candidates. Their Normfinder stability values calculated across the primary tumor and metastases subgroup indicate that miR-29a-3p can be considered as the strongest housekeeper in a cohort with mainly samples from primary tumors, whereas miR-16-5p might perform better in a metastatic sample enriched cohort.

【 授权许可】

Unknown