ECEESPE2025 ePoster Presentations Adrenal and Cardiovascular Endocrinology (170 abstracts)
Pitfalls of circulating miRNA-based biomarker studies in adrenocortical tumors
Bálint Vékony 1,2 , Gábor Nyirő 1,2,3 , Henriett Butz 3,4,5 , Bálint Szeredás 1,2 , Viktória Tóth 6 , Peter Ferdinandy 6 , Attila Patócs 3,4,5 & Peter Igaz 1,2
1Semmelweis University, Department of Internal Medicine and Oncology, Budapest, Hungary; 2Semmelweis University, Department of Endocrinology, Budapest, Hungary; 3Semmelweis University, Department of Laboratory Medicine, Budapest, Hungary; 4HUN-REN-SU, Hereditary Tumours Research Group, Budapest, Hungary; 5National Institute of Oncology, Department of Molecular Genetics and the National Tumour Biology Laboratory, Budapest, Hungary; 6Semmelweis University, Department of Pharmacology and Pharmacotherapy, Budapest, Hungary
JOINT1657
Introduction: Differentiating between benign and malignant adrenocortical tumors has major clinical relevance. Recent studies have highlighted the potential of circulating microRNAs (miRNAs) as biomarkers for malignant adrenocortical carcinoma (ACC). However, there are many difficulties with their use, mainly standardization.
Aims: Our aim was to compare the interchangeability of real-time quantitative polymerase chain reaction (qPCR) and digital PCR (dPCR) in measuring circulating miRNAs, and to investigate whether the use of K2 or K3 EDTA anticoagulants may influence the results.
Methods: Peripheral blood samples were taken simultaneously into K2 and K3 EDTA collection tubes, from 20 individuals. After sample procession, three miRNAs associated with ACC (hsa-miR-483-5p, -210-3p, -21-5p) as well as two controls (-miR-16-5p, cel-miR-39-3p) were analyzed utilizing RT-qPCR and dPCR. Data obtained were compared by the following statistical methods: Spearman’s rank correlation, paired t tests and Bland-Altman analysis.
Results: qPCR and dPCR results show a significant correlation (p values between 0.0072 and 0.049) in K2-EDTA samples when comparing ΔCt values and copy numbers. However, proportional biases relating to low and high miRNA expression were observed between the two methods. In qPCR measurements, K3-EDTA results showed higher standard deviations (average SD for K2 samples was 0.91 while for K3 samples 1.1). When comparing raw Ct values, only miR-483-5p was found to be significantly different, while in case of ΔCt values, every miRNA except miR-483-5p was significantly different. dPCR results were not affected by the different anticoagulants.
Conclusions: dPCR and qPCR are not easily interchangeable, especially for rare or abundant miRNAs, making cross-validation difficult. The choice of EDTA could potentially influence qPCR results, highlighting the need for standardized protocols.
Volume 110
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Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
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