Petrova Tatiana, Kalinina Olga, Aquino Arthur, Grigoryev Evgeniy, Dubashynskaya Natallia V, Zubkova Kseniya, Kostareva Anna, Golovkin Alexey
Almazov National Medical Research Centre, 197341 St. Petersburg, Russia.
St. Petersburg State University, Research Park, 199034 St. Petersburg, Russia.
Noncoding RNA. 2024 Feb 15;10(1):15. doi: 10.3390/ncrna10010015.
There are many articles on the quantitative analysis of miRNAs contained in a population of EVs of different sizes under various physiological and pathological conditions. For such analysis, it is important to correctly quantify the miRNA contents of EVs. It should be considered that quantification is skewed depending on the isolation protocol, and different miRNAs are degraded by nucleases with different efficiencies. In addition, it is important to consider the contribution of miRNAs coprecipitating with the EVs population, because the amount of miRNAs in the EVs population under study is skewed without appropriate enzymatic treatment. By studying a population of EVs from the blood plasma of healthy donors, we found that the absolute amount of miRNA inside the vesicles is commensurate with the amount of the same type of miRNA adhered to the outside of the EVs. The inside/outside ratio ranged from 1.02 to 2.64 for different investigated miRNAs. According to our results, we propose the hypothesis that high occupancy of miRNAs on the outer surface of EVs influence on the transporting RNA repertoire no less than the inner cargo received from the host cell.
有许多关于在各种生理和病理条件下对不同大小的细胞外囊泡群体中所含miRNA进行定量分析的文章。对于此类分析,正确量化细胞外囊泡的miRNA含量很重要。应该考虑到,根据分离方案,定量会出现偏差,并且不同的miRNA会被核酸酶以不同的效率降解。此外,考虑与细胞外囊泡群体共沉淀的miRNA的贡献也很重要,因为在没有适当酶处理的情况下,所研究的细胞外囊泡群体中的miRNA量会出现偏差。通过研究健康供体血浆中的细胞外囊泡群体,我们发现囊泡内miRNA的绝对量与附着在细胞外囊泡外部的同类型miRNA的量相当。对于不同研究的miRNA,内部/外部比率在1.02至2.64之间。根据我们的结果,我们提出一个假设,即细胞外囊泡外表面上miRNA的高占有率对转运RNA库的影响不亚于从宿主细胞接收的内部货物。