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Increased levels of circulating cell-free double-stranded nucleic acids in the plasma of glioblastoma patients.
Rackles, Elisabeth; Zaccheroni, Elena; Lopez, Patricia Hernandez; Faletti, Stefania; Bene, Massimiliano Del; DiMeco, Francesco; Pelicci, Giuliana; Falcon-Perez, Juan M.
Afiliación
  • Rackles E; Center for Cooperative Research in Biosciences (CIC bioGUNE) Basque Research and Technology Alliance (BRTA) Derio Spain.
  • Zaccheroni E; Department of Experimental Oncology European Institute of Oncology (IEO) IRCCS Milan Italy.
  • Lopez PH; Center for Cooperative Research in Biosciences (CIC bioGUNE) Basque Research and Technology Alliance (BRTA) Derio Spain.
  • Faletti S; Department of Experimental Oncology European Institute of Oncology (IEO) IRCCS Milan Italy.
  • Bene MD; Department of Neurosurgery Fondazione IRCCS Istituto Neurologico Carlo Besta Milan Italy.
  • DiMeco F; Department of Neurosurgery Fondazione IRCCS Istituto Neurologico Carlo Besta Milan Italy.
  • Pelicci G; Department of Pathophysiology and Transplantation University of Milan Milan Italy.
  • Falcon-Perez JM; Department of Neurological Surgery Johns Hopkins Medical School Baltimore Maryland USA.
J Extracell Biol ; 3(8): e168, 2024 Aug.
Article en En | MEDLINE | ID: mdl-39100684
ABSTRACT
Circulating cell-free nucleic acids are considered a promising source of biomarkers for diseases and cancer. Liquid biopsy biomarkers for brain tumours represent a major, still unmet, clinical need. In plasma, nucleic acids can be free or be associated with extracellular vesicles (EVs). Here we report an easy and reproducible method to analyse cell-free nucleic acids in plasma and EVs by conventional flow cytometry easy to translate into the clinics. Nucleic acids associated with the EVs or present in plasma samples are stained by Pyronin Y, which is a fluorescent dye that is preferably binding double-stranded nucleic acids. Fluorescent staining of EVs isolated from cell-conditioned media is suitable for DNA and RNA detection by flow cytometry. The nucleic acids are partially protected from degradation by the EVs' membrane. Additionally, DNA and RNA can be stained in plasma samples and plasma-derived EVs. Remarkably, analysis of plasma from patients and healthy individuals reveals a difference in their nucleic acid profiles. Taken together, our results indicate that the proposed methodology, which is based on conventional direct flow cytometry, is a promising easy tool for plasma nucleic acid analysis.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Extracell Biol Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: J Extracell Biol Año: 2024 Tipo del documento: Article Pais de publicación: Estados Unidos