Tag Archives: carcinogenesis

Extracellular vesicles produced by mouse breast adenocarcinoma 4T1 cells with up- or down-regulation of adaptor protein Ruk/CIN85 differentially modulate the biological properties of 4T1 WT cells

A. Yu. Zhyvolozhnyi1,2*, I. R. Horak1, D. S. Geraschenko1, M. O. Gomozkova3,
O. O. Hudkova1, S. J. Vainio2, A. A. Samoylenko2, L. B. Drobot1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
2Faculty of Biochemistry and Molecular Medicine, University of Oulu, Finland;
3Brigham Young University-Idaho, Rexburg, USA;
*e-mail: ppndl2@gmail.com

Received: 02 November 2021; Accepted: 12 November 2021

Extracellular vesicles (EVs) are secreted by most cell types under both physiological and pathological conditions and were proposed to be actively involved in intercellular communication. The mode of EVs action is dependent on their cargos composition. EVs  play an important role in tumor initiation, recurrence, metastasis and therapeutic resistance. EVs marker proteins Alix and Tsg101 and cortactin are the binding partners of adaptor protein Ruk/CIN85. The present study aims to analyze the regulatory effects of EVs produced by 4T1 cells with overexpression (RukUp) or down-regulation (RukDown) of adaptor protein Ruk/CIN85 on proliferation rate, migration and invasion activity of parental 4T1 WT cells. EVs from conditioned medium of 4T1 RukUp or RukDown cells were isolated by differential centrifugation followed by further purification using Exo-spin™ kit (Cell Guidance Systems). The number and size of EVs were characterized by NTA (Malvern Panalytical NanoSight NM300) instrument. The content of marker proteins and Ruk/CIN85 in isolated EVs was analyzed by Western-blotting. The viability, migration and invasion activity of 4T1 WT cells were studied using MTT-test, scratch-test and Boyden chamber assay, respectively. It was demonstrated for the first time that adaptor protein Ruk/CIN85 is a constitutive component of  EVs produced by 4T1 cells. It was also shown that EVs produced by 4T1 cells with different levels of Ruk/CIN85 expression are characterized by a specific profile of the content of its multiple molecular forms. It turned out that the ability of EVs to modulate the proliferative activity, motility and invasiveness of 4T1 WT cells was tightly correlated with the biological properties of 4T1 cells that produce EVs (highly aggressive 4T1 RukUp cells or weakly invasive 4T1 RukDown cells). Our data suggest that adaptor protein Ruk/CIN85 is not only a constitutive component of cargos composition of EVs produced by tumor cells but, depending on its content in EVs, plays an active role in the control of carcinogenesis.

Scientific and practical activity of the Laboratory of Cell Signaling Mechanisms of the Palladin Institute of Biochemistry of NAS of Ukraine

R. P. Vynogradova, I. Yu. Chernysh, V. M. Danilova

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: valdan@biochem.kiev.ua

The article is devoted to the analysis of the scientific and practical activity of the laboratory of the signaling mechanisms of the cells of the Palladin Institute of Biochemistry, NAS of Ukraine, in the context of the history of its development. The most important results of studies of the mechanisms controlling proliferation, migration and invasion of tumor cells, which testify to the important role of the Ruk/CIN85 adapter protein in carcinogenesis, are presented. These studies are a priority and can serve as an experimental basis for the development of new generation pharmacological agents, the targets for which can be key centers for the organization of signaling systems of the cell – adapter and scaffold proteins.

Reactive oxygen species in signal transduction

L. B. Drobot1, A. A. Samoylenko1, A. V. Vorotnikov2, P. A. Tyurin-Kuzmin2,
A. V. Bazalii1, T. Kietzmann3, V. A. Tkachuk2, S. V. Komisarenko1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: drobot@biochem.kiev.ua
2Lomonosov Moscow State University, Faculty of Basic Medicine, Russia;
3Department of Biochemistry and Biocenter Oulu, University of Oulu, Oulu, Finland;

Reactive oxygen species (ROS) are products of incomplete reduction of oxygen both nonradicals and radicals that function as mediators of redox signaling and oxidative stress depending on their levels in different­ subcellular compartments. Up to date, a huge body of data are accumulated, which supports a role of ROS as “second messengers” in intracellular signaling cascades that control cell growth, proliferation, apoptosis as well as migration and invasion. The current review summarizes data regarding ROS-dependent regulation of signaling­ networks components including MAPK, PI3K/Akt, PKC, NF-κB, Nrf2, FoxO and HIF-1α, and role of ROS in tumorigenesis.

Effect of dihydropyrrol and maleimide derivatives on the state of the liver and colon in normal rats and those with colorectal carcinogenesis induced by dimethylhydrazine

H. M. Kuznietsova, O. V. Lynchak, M. O. Danylov, I. P. Kotlyar, V. K. Rybalchenko

Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: gala_kuznetsova@rambler.ru

No liver and colon alterations in rats, caused by cytostatic compounds 5-amino-4-(1,3-benzothyazol-2-yl)-1-(3-methoxyphenyl)-1,2-dihydro-3Н-pyrrol-3-one (D1) and 1-(4-Cl-benzyl)-3-Cl-4-(CF3-phenylamino)-1H-pyrrol-2,5-dione (MI-1) when administered over a long time were found, as evidenced by the histopathological data and the data of activity of transaminases, alkaline phosphatase and lactate dehydrogenase in the blood serum. D1 and MI-1 in vivo decrease the total area of DMH-induced colon tumors in rats by 46-60%. Furthermore, D1 and MI-1 partially protect the liver and colon mucosa from toxic effects caused by 1,2-dimethylhydrazine (DMH) reducing DNA oxidative modifications, as evidenced by urine 8-hydroxydeoxyguanosine level. The effects of both compounds are similar, but MI-1 is less toxic for the liver and colon of intact animals possessing more pronounced antitumor activity and protective properties in the setting of chemically induced carcinogenesis.