MiR-26a-5p as a Reference to Normalize MicroRNA qRT-PCR Levels in Plasma Exosomes of Pediatric Hematological Malignancies

Cells

Enrico Gaffo¹  Stefania Bortoluzzi¹  Federica Lovisa²  Piero Di Battista²  Anna Tosato²  Alessandra Biffi²  CarlottaC. Damanti²  Lara Mussolin²  Anna Garbin²  Ilaria Gallingani²  Marta Pillon³  Elisa Carraro³  Caterina Elia⁴  Maurizio Mascarin⁴ 

[1] Department of Molecular Medicine, Padova University, 35121 Padova, Italy;Maternal and Child Health Department, Padova University, 35128 Padova, Italy;Pediatric Hematology, Oncology and Stem Cell Transplant Division, Padova University Hospital, 35128 Padova, Italy;Pediatric Radiotherapy Unit, Centro di Riferimento Oncologico (CRO) di Aviano, IRCCS, 33081 Aviano, Italy;

关键词: circulating microRNAs;    exosomes;    qRT-PCR;    normalization;    reference genes;    hematological malignancies;   

DOI  :  10.3390/cells10010101

来源: DOAJ

【 摘 要 】

Plasma exosomal microRNAs (miRNAs) are considered as valid circulating biomarkers for cancer diagnosis and prognosis. Quantitative real-time polymerase chain reaction (qRT-PCR), the most commonly used technique to assess circulating miRNA levels, requires a normalization step involving uniformly expressed endogenous miRNAs. However, there is still no consensus on reference miRNAs for plasma exosomal miRNA abundance normalization. In this study, we identified a panel of miRNAs with stable abundance by analyzing public plasma exosome RNA-seq data and selected miR-486-5p, miR-26a-5p, miR-423-5p and miR191-5p as candidate normalizers. Next, we tested the abundance variation of these miRNAs by qRT-PCR in plasma exosomes of healthy donors and pediatric patients with anaplastic large cell lymphoma, Burkitt lymphoma, Hodgkin lymphoma and mature B-cell acute lymphoblastic leukemia. MiR-486-5p and miR-26a-5p showed the most stable levels, both between healthy controls and patients and among the malignancies analyzed. In light of previous reports on miRNA stability in different exosome isolation methods, our data indicated that miR-26a-5p is a bona fide reference miRNA for qRT-PCR normalization to evaluate miRNA abundance from circulating plasma exosomes in studies of hematological malignancies.

【 授权许可】

Unknown