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Biomolecules. 2020 Jan 16;10(1). pii: E150. doi: 10.3390/biom10010150.

Identification of miRNAs Enriched in Extracellular Vesicles Derived from Serum Samples of Breast Cancer Patients.

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Department of Genetics, Federal University of Paraná, Curitiba 81531-980, Brazil.
Department of Cell and Molecular Biology, Federal University of Paraná, Curitiba 81531-980, Brazil.
Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba 81531-980, Brazil.
Carlos Chagas Institute - Fiocruz-Paraná, Curitiba 81350-010, Brazil.
Positivo University Medical School, Curitiba 81280-330, Brazil.
Nossa Senhora das Graças Hospital Breast Unit, Curitiba 80810-040, Brazil.
Department of Biomedical Data Sciences, Molecular Epidemiology, Leiden University Medical Center, 2300 R Leiden, The Netherlands.
Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC 20057, USA.
Research Institute Pelé Pequeno Príncipe, Faculdades Pequeno Príncipe, Curitiba 80230-020, Brazil.


MicroRNAs derived from extracellular vesicles (EV-miRNAs) are circulating miRNAs considered as potential new diagnostic markers for cancer that can be easily detected in liquid biopsies. In this study, we performed RNA sequencing analysis as a screening strategy to identify EV-miRNAs derived from serum of clinically well-annotated breast cancer (BC) patients from the south of Brazil. EVs from three groups of samples (healthy controls (CT), luminal A (LA), and triple-negative (TNBC)) were isolated from serum using a precipitation method and analyzed by RNA-seq (screening phase). Subsequently, four EV-miRNAs (miR-142-5p, miR-150-5p, miR-320a, and miR-4433b-5p) were selected to be quantified by quantitative real-time PCR (RT-qPCR) in individual samples (test phase). A panel composed of miR-142-5p, miR-320a, and miR-4433b-5p distinguished BC patients from CT with an area under the curve (AUC) of 0.8387 (93.33% sensitivity, 68.75% specificity). The combination of miR-142-5p and miR-320a distinguished LA patients from CT with an AUC of 0.9410 (100% sensitivity, 93.80% specificity). Interestingly, decreased expression of miR-142-5p and miR-150-5p were significantly associated with more advanced tumor grades (grade III), while the decreased expression of miR-142-5p and miR-320a was associated with a larger tumor size. These results provide insights into the potential application of EVs-miRNAs from serum as novel specific markers for early diagnosis of BC.


RNA-seq; circulating microRNAs; extracellular vesicles; liquid biopsy; miRNA

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