دورية أكاديمية
أعرض في EDS

Comparison of Methods for MicroRNA Isolation from Extracellular Vesicles Obtained from Ascitic Fluids.

التفاصيل البيبلوغرافية
العنوان: Comparison of Methods for MicroRNA Isolation from Extracellular Vesicles Obtained from Ascitic Fluids.
المؤلفون: Skryabin GO; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Vinokurova SV; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Elkina NV; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Denisova DA; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Beliaeva AA; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia.; Faculty of Biology, Lomonosov Moscow State University, Moscow, 111234, Russia., Zhordania KI; Institute of Clinical Oncology, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Bagrov DV; Faculty of Biology, Lomonosov Moscow State University, Moscow, 111234, Russia., Enikeev AD; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Galetsky SA; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Komelkov AV; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia. komelkov@gmail.com., Krasnoshekova GI; Institute of Clinical Oncology, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia., Tchevkina EM; Institute of Carcinogenesis, Blokhin National Medical Research Center of Oncology, Moscow, 115478, Russia.
المصدر: Biochemistry. Biokhimiia [Biochemistry (Mosc)] 2022 Nov; Vol. 87 (11), pp. 1354-1366.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: MAIK Nauka/Interperiodica Country of Publication: United States NLM ID: 0376536 Publication Model: Print Cited Medium: Internet ISSN: 1608-3040 (Electronic) Linking ISSN: 00062979 NLM ISO Abbreviation: Biochemistry (Mosc) Subsets: MEDLINE
أسماء مطبوعة: Publication: <2007->: Moscow : MAIK Nauka/Interperiodica
Original Publication: New York, Consultants Bureau [etc.]
مواضيع طبية MeSH: MicroRNAs*/metabolism , Extracellular Vesicles*/metabolism , Exosomes*/metabolism, Ascitic Fluid/metabolism
مستخلص: Secreted extracellular vesicles (EVs) contain active biomolecules, including miRNAs, composition of which reflects epigenetic changes occurring in cells during pathological processes, in particular, malignant transformation. The accumulated pool of data on the role of EVs in carcinogenesis has stimulated investigations of the EV-derived cancer markers. The most important factor limiting development of this scientific direction is lack of "gold standards" both for methods of EV isolation from biological fluids and for analyzing their molecular content, including composition of miRNAs. Here we first examined efficacy of various methods for small RNA isolation from EVs contained in ascitic fluid for subsequent miRNA analysis. Comparison of different commercial kits showed advantages of the methods based on phenol-chloroform extraction: Total Exosome RNA & Protein Isolation Kit and miRNeasy Serum/Plasma Kit. Analysis of the small RNA transcriptome showed presence of various classes of molecules in the EVs, among which proportion of miRNAs averaged 6% and reaching 10% with the Total Exosome RNA & Protein Isolation Kit. The PureLink miRNA Isolation Kit demonstrated the lowest efficiency. The miRNeasy Advanced Serum/Plasma Kit showed the highest concentration of the small RNA fraction, miRNA proportion of which, however, did not exceed that obtained with the miRNeasy Serum/Plasma Kit and Total Exosome RNA & Protein Isolation Kit. Moreover, RT-PCR analysis of the individual molecules showed lower levels of each of investigated miRNAs (miR-1246, miR-200b-5p, miR-200c-3p, and miR-23a-3p) when using the miRNeasy Advanced Serum/Plasma Kit. In conclusion, Total Exosome RNA & Protein Isolation Kit and miRNeasy Serum/Plasma Kit can be considered as optimal kits in terms of performance based on combination of the studied characteristics, including small RNA concentration, percentage of microRNA according to bioanalyzer and sequencing results, and levels of individual miRNAs detected by RT-PCR.
فهرسة مساهمة: Keywords: PureLink miRNA Isolation Kit; Total Exosome RNA & Protein Isolation Kit; exosomes; extracellular vesicles; isolation methods; miRNA; miRNeasy Advanced Serum/Plasma Kit; miRNeasy Serum/Plasma Kit
المشرفين على المادة: 0 (MicroRNAs)
تواريخ الأحداث: Date Created: 20221212 Date Completed: 20221214 Latest Revision: 20221221
رمز التحديث: 20240628
DOI: 10.1134/S0006297922110141
PMID: 36509726
قاعدة البيانات: MEDLINE
الوصف
تدمد:1608-3040
DOI:10.1134/S0006297922110141