BioProject

Circulating microRNAs (miRNAs) may be able to serve as prognostic biomarkers. More...

Circulating microRNAs (miRNAs) may be able to serve as prognostic biomarkers. However, identifying the accurate screening method/s for miRNA detection/quantification from serum is crucial for any further downstream analysis. Here, we compared two high-throughput methods, small RNA sequencing (sRNA-seq) and TaqMan Low-Density Array (TLDA), to quantify miRNAs at baseline (time of primary surgery) in the serum from ER+ BC patients for whom the long-term clinical follow-up information (10 years) and subsequent metastasis status are known. For this study, 48 serum samples were selected based on the outcome of breast cancer patients, and therefore sub-divided in: patients with no recurrence within a 10-years period (Cohort 1, n=16), patients with early recurrence within 5 years (Cohort 2, <5 Years, n=16), or patients with late recurrence after 5 years up to 10 years (Cohort 3, >5 Years, n=16). For each serum sample, the miRNome analysis was performed by Next Generation Sequencing (NGS) using 20 µl of the 45 µl total RNA extracted from 200 µl serum by the miRCURY Biofluids RNA Isolation Kit. The NGS libraries were constructed and sequenced at the ProfileXpert platform (www.profilexpert.fr). The data was quality checked by assigning Q-scores after the intensity correction and base calling. The raw data were treated with the NCS 2.0.2 and RTA 2.4.11 software’s to generate the Base Calling File. The Bcl2factq v2.17.1.14 was used to demultiplexing using 1 mismatch. Less...