Preview

Bulletin of Siberian Medicine

Advanced search

DISRUPTION OF EXPRESSION OF MRNA HSP27 AND UBIQUITIN AS A MECHANISM OF ESCAPING OF JURKAT LINE TUMOR CELLS FROM APOTOSIS

https://doi.org/10.20538/1682-0363-2015-1-66-72

Abstract

The research objective is to establish the link between heat shock protein 27 and ubiquitin mRNA expression as well as Jukart tumor cell apoptosis.
The method of flow cytofluorometry has been used to evaluate apoptosis realization using FITC-labeled annexin V and propidium iodide along with the amount of reactive oxygen species. Spectrofluorimetry has been applied to register the caspase-3 activity. The content of hydroxyl radicals has been determined by spectrophotometry. The level of ubiquitin and heat shock protein 27 mRNA expression has been identified using real-time PCR. Intact Jukart tumor cells and blood lymphocytes of healthy donors served the material for the research.
Following the carried out research it has been found out that the fall in the amount of annexin V positive cells and the reduced caspase-3 activity were accompanied by the rise in the content of hydroxyl radicals and reactive oxygen species against the backdrop of the increased heat shock protein 27 and ubiquitin mRNA expression in Jukart tumor cells.

About the Authors

O. L. Nosareva
Сибирский государственный медицинский университет, Томск
Russian Federation
Siberian State Medical University, Tomsk


Ye. A. Stepovaya
Siberian State Medical University, Tomsk
Russian Federation


N. V. Ryazantseva
Siberian State Medical University, Tomsk
Russian Federation


Ye. V. Zakirova
Siberian State Medical University, Tomsk
Russian Federation


I. O. Mazunin
Immanuel Kant Baltic Federal University, Kaliningrad
Russian Federation


L. S. Litvinova
Immanuel Kant Baltic Federal University, Kaliningrad
Russian Federation


N. A. Sokhonevich
Immanuel Kant Baltic Federal University, Kaliningrad
Russian Federation


O. N. Vesnina
Siberian State Medical University, Tomsk
Russian Federation


Ye. V. Shakhristova
Siberian State Medical University, Tomsk
Russian Federation


References

1. Ryazantseva N.V., Kaigorodova Ye.V., Maroshkina A.N. et al. The effects of proteins of thermal shock modulating apoptosis: influence of the chaperon of Hsp27 on proteins of Bcl2 family in tumor cells of the Jurkat line. Voprosy onkologii – Problems in Oncology, 2012, vol. 58, no. 4, pp. 541–544 (in Russian).

2. Negroni L., Samson M., Guigonis J.M. et al. Treatment of colon cancer cells using the cytosine deaminase/5fluorocytosine suicide system induces apoptosis, modulation of the proteome, and Hsp90beta phosphorylation. Cancer Ther., 2007, vol. 6, no. 10, pp. 2747–2756.

3. Saveleva O.E., Anishchenko E.A., Novitsky V.V., Riazantseva N.V. The role of transcription factors in cytokine-mediated apoptosis of lymphocytes. International Journal of Biology, 2012, vol. 4, no. 1, pp. 129–137.

4. Zenkov N.K., Men'shhikova E.B., Tkachev V.O. Some principles and mechanisms of redoks-regulation. Kislorod i antioksidanty – Oxygen and Antioxidants, 2009, no. 1, pp. 3–64 (in Russian).

5. Ramakrishna S., Suresh B., Baek K.-H. The role of deubiquitinating enzymes in apoptosis. Cell. Mol. Life Sci., 2011, no. 68, pp. 15–26.

6. Cimoha A.S. Proteasoma: participation in cellular processes. Cytology, 2010, vol. 52, no. 4, pp. 271–300.

7. Raab S., Drechsel G., Zarepour M. et al. Identification of a novel E3 ubiquitin ligase that is required for suppression of premature senescence in Arabidopsis. Plant J., 2009, vol. 59, no. 1, pp. 39–51.

8. Mel'nikov E.E., Rotanova T.V. Molecular chaperons. Boorganicheskaya khimiya – Bioorganic Chemistry, 2010, vol. 36, no. 1, pp. 5–14 (in Russian).

9. Ryazantseva N.V., Kaigorodova Ye.V., Novitsky V.V. et al. Molecular participants of a receptor way of regulation of apoptosis in tumoral and normal lymphocytes in the co nditions of inhibition of protein of thermal shock of 90 in vitro. Sibirskiy onkologicheskiy zhurnal – Siberian Journal of Oncology, 2011, vol. 2, no. 44, pp. 52–56 (in Russian).

10. Richter K., Haslbeck M., Buchner J. The heat shock response: life on the verge of death. Mol. Cell., 2010, vol. 40, no. 2, pp. 253–266.

11. Bignold L.P., Ferrante A. Mechanism of separation of polymorphonuclear leukocytes from whole blood by the onestep Hypaque-Ficoll method. J. Immunol. Methods, 1987, vol. 96, no. 1, pp. 29–33.

12. Ulmer A.J., Flad H.D. Discontinuous density gradient separation of human mononuclear leukocytes using Percoll as gradient medium. J. Immunol. Methods, 1979, vol. 30, no. 1, pp. 1–10.

13. Cohen G.M. Caspases: the executioners of apoptosis. Biochem. J., 1997, vol. 326, pp. 1–16.

14. Nicholson D.W. Caspase structure, proteolytic substrates, and function during apoptotic cell death. Cell Death and Differentiation, 1999, vol. 6, pp. 1028–1042.

15. Bass D.A., Parce J.W., Dechatelet L.R. et al. Flow cytometric studies of oxidative product formation by neutrophils: A graded response to membrane stimulation. J. Immunol., 1983, vol. 130, pp. 1910–1917.

16. Thom S.R., Elbuken M.E. Oxygen-dependent antagonism of lipid perodixation. Free Radical Biol. Med., 1991, vol. 10, no. 6, pp. 413–426.

17. Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt. Biochem., 1976, vol. 7, no. 1, 2, pp. 248–254.

18. Schmittgen T.D., Livak K.J. Analyzing real-time PCR data by the comparative C (T) method. Nat. Protoc., 2008, vol. 3, no. 6, pp. 1101–1108.

19. Glants S. Medicobiological statistics. Moscow, Practice Publ., 1999. 459 p. (in Russian).

20. Beere H.M. “The stress of dying”: the role of heat shock proteins in the regulation of apoptosis. Cell Sci., 2004, vol. 117, no. 13, pp. 2641–2651.

21. Dubinina E.E., Pustygina A.V. Oxidizing modification of proteins, its role at pathological states. Ukrainskiy biokhimicheskiy zhurnal – The Ukrainian Biochemical Magazine, 2008, vol. 80, no. 6, pp. 5–18 (in Russian).

22. Men'shhikova E.B., Zenkov N.K., Lankin V.Z. et al. Oxidizing stress: Pathological states and diseases. Novosibirsk, ARTA Publ., 2008. 284 p. (in Russian).

23. Stetler R.A. HSP27: mechanisms of cellular protection against neuronal injury. Curr. Mol. Med., 2009, vol. 9, no. 7, pp. 863–872.


Review

For citations:


Nosareva O.L., Stepovaya Ye.A., Ryazantseva N.V., Zakirova Ye.V., Mazunin I.O., Litvinova L.S., Sokhonevich N.A., Vesnina O.N., Shakhristova Ye.V. DISRUPTION OF EXPRESSION OF MRNA HSP27 AND UBIQUITIN AS A MECHANISM OF ESCAPING OF JURKAT LINE TUMOR CELLS FROM APOTOSIS. Bulletin of Siberian Medicine. 2015;14(1):66-72. (In Russ.) https://doi.org/10.20538/1682-0363-2015-1-66-72

Views: 706


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


ISSN 1682-0363 (Print)
ISSN 1819-3684 (Online)