Identification of stably expressed microRNAs in plasma from high-grade serous ovarian carcinoma and benign tumor patients

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Background: Ovarian cancer is a lethal gynecological cancer and no reliable minimally invasive early diagnosis tools exist. High grade serous ovarian carcinoma (HGSOC) is often diagnosed at advanced stages, resulting in poorer outcome than those diagnosed in early stage. Circulating microRNAs have been investigated for their biomarker potential. However, due to lack of standardization methods for microRNA detection, there is no consensus, which microRNAs should be used as stable endogenous controls. We aimed to identify microRNAs that are stably expressed in plasma of HGSOC and benign ovarian tumor patients. Methods and results: We isolated RNA from plasma samples of 60 HGSOC and 48 benign patients. RT-qPCR was accomplished with a custom panel covering 40 microRNAs and 8 controls. Stability analysis was performed using five algorithms: Normfinder, geNorm, Delta-Ct, BestKeeper and RefFinder using an R-package; RefSeeker developed by our study group [1]. Among 41 analyzed RNAs, 13 were present in all samples and eligible for stability analysis. Differences between stability rankings were observed across algorithms. In HGSOC samples, hsa-miR-126-3p and hsa-miR-23a-3p were identified as the two most stable miRNAs. In benign samples, hsa-miR-191-5p and hsa-miR-27a-3p were most stable. In the combined HGSOC and benign group, hsa-miR-23a-3p and hsa-miR-27a-3p were identified by both the RefFinder and Normfinder analysis as the most stable miRNAs. Conclusions: Consensus regarding normalization approaches in microRNA studies is needed. The choice of endogenous microRNAs used for normalization depends on the histological content of the cohort. Furthermore, normalization also depends on the algorithms used for stability analysis.

OriginalsprogEngelsk
TidsskriftMolecular Biology Reports
Vol/bind50
Udgave nummer12
Sider (fra-til)10235-10247
Antal sider13
ISSN0301-4851
DOI
StatusUdgivet - 2023

Bibliografisk note

Funding Information:
We are thankful to the Danish CancerBiobank (Bio- and GenomeBank, Denmark - RBGB) and the Danish Gynecologic Cancer Database (DGCD) for making samples and data available for use in this study. This work was founded by: The Mermaid Foundation, URL: http://www.mermaidprojektet.dk/ (PHDP, JLJ, CKH and EVH received the funding), Danish Cancer Research Foundation: URL: http://www.dansk-kraeftforsknings-fond.dk/ (EVH received the funding), and Herlev Hospital Research Council, URL: https://www.herlevhospital.dk/forskning/ (EVH received the funding).

Funding Information:
We are thankful to the Danish CancerBiobank (Bio- and GenomeBank, Denmark - RBGB) and the Danish Gynecologic Cancer Database (DGCD) for making samples and data available for use in this study. This work was founded by: The Mermaid Foundation, URL: http://www.mermaidprojektet.dk/ (PHDP, JLJ, CKH and EVH received the funding), Danish Cancer Research Foundation: URL:http://www.dansk-kraeftforsknings-fond.dk/ (EVH received the funding), and Herlev Hospital Research Council, URL: https://www.herlevhospital.dk/forskning/ (EVH received the funding).

Funding Information:
This study was funded by the The Mermaid Foundation, URL: http://www.mermaidprojektet.dk/ (PHDP, JLJ, CKH and EVH received the funding), Danish Cancer Research Foundation, URL: http://www.dansk-kraeftforsknings-fond.dk/ (EVH received the funding) as well as Herlev Hospital Research Council https://www.herlevhospital.dk/forskning/ (EVH received the funding).

Publisher Copyright:
© 2023, The Author(s).

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