University of Oulu

Zhyvolozhnyi, A. Yu., Horak, I. R., Skaterna, T. D., Khudiakova, O. V., Vainio, S. J., Samoylenko, A. A., & Drobot, L. B. (2021). Composition of EVs markers under normoxic and hypoxic conditions depends on the expression level of adaptor protein Ruk/CIN85 in mouse renal carcinoma Renca cells. Biopolymers and Cell, 37(5), 325–334.

Composition of EVs markers under normoxic and hypoxic conditions depends on the expression level of adaptor protein Ruk/CIN85 in mouse renal carcinoma Renca cells

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Author: Zhyvolozhnyi, A. Yu.1,2; Horak, I. R.2; Skaterna, T. D.2;
Organizations: 1University of Oulu Linnanmaa, PO Box 3000, FIN-90014 Oulu, Finland
2Palladin Institute of Biochemistry, NAS of Ukraine 9, Leontovycha Str., Kyiv, Ukraine, 01601
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 0.6 MB)
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Language: English
Published: National Academy of Sciences of Ukraine, 2021
Publish Date: 2022-03-25


Aim: To isolate and characterize extracellular vesicles (EVs) produced by mouse renal carcinoma Renca cells with different expression levels of the adaptor protein Ruk/CIN85 under normoxia and hypoxia conditions.

Methods: The density gradient centrifugation was used to isolate EVs from the conditioned medium of Renca cells cultured under normoxia and hypoxia conditions. Further characterization of EVs was performed by using nanoparticle tracking analysis (NTA), electron microscopy and Western Blot analysis.

Results: Significant differences in average particle size between EVs produced by sublines studied under experimental conditions were not found. At the same time, concentration of particles produced by Ruk/CIN85 overexpressing cells turned out to be an order of magnitude higher in hypoxia in comparison to normoxia conditions. It was shown that under normoxia conditions the content of both Ruk/CIN85 and EVs’ markers Alix and CD81 was increased in vesicles produced by Renca cells with Ruk/CIN85 overexpression in comparison with those from control mock-transfected cells. Under hypoxia conditions, the content of studied proteins decreased by more than two orders of magnitude in EVs secreted by Renca cells with up-regulation of adaptor protein whereas the content of Ruk/CIN85 and CD81 increased in EVs from mock-transfected cells.

Conclusions: It has been demonstrated that the adaptor protein Ruk/CIN85 is a novel component of EVs produced by tumor cells that may play a role in the control of EV composition under normoxia and hypoxia.

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Series: Biopolymers and cell
ISSN: 0233-7657
ISSN-E: 1993-6842
ISSN-L: 0233-7657
Volume: 37
Issue: 5
Pages: 325 - 334
DOI: 10.7124/bc.000A5E
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
3122 Cancers
Funding: The study was supported by the EDUFI (Finnish National Agency for Education) fellowships to A. Yu. Zhyvolozhnyi.
Copyright information: © 2021 A. Yu. Zhyvolozhnyi et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Biopolymers and Cell. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.