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VIֲ̲ߠ Ҳ ϲҲ  ܠϠ ò ʠǠ (2124ߠ2010,.²) ...

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̲ͲΠ²ȠȠȠ

²ɠֲɠͲҠͲ

VIֲ̲ߠ

Ҳ ϲҲ 

ܠϠ

ò ʠǠ

(2124ߠ2010,.²)

² 2010

MINISTRYOFEDUCATIONANDSCIENCEOFUKRAINE

IVANFRANKONATIONALUNIVERSITYOFLVIV

VIINTERNATIONALSCIENTIFICCONFERENCE

FORSTUDENTSANDPHDSTUDENTS YOUTHANDPROGRESSOF BIOLOGY ABSTRACTSBOOK (SEPTEMBER2124,2010,LVIV) LVIV2010 581.1:57 㳿: VI ̳ (21-24 2010 , . ). , 2010. 240 .

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, - "-", (The Friends of Lviv University, Inc, USA).

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Youth and Progress of Biology: abstracts book of the VI International Scientific Conference of Students and PhD Students (September 21-24, 2010, Lviv). Lviv, 2010. 240 p.

Abstracts book contains the results of scientific work of students and PhD students of Ukraine and foreign countries. The book is reckoned on the scientists, PhD students and students, which are working in the field of biology and biomedicine.

The authors are responsible for the trustworthiness of scientific results and for the text.

The organizers of the conference thank the Rector's Office of Ivan Franko National University of Lviv, the Ukrainian-American beneficial fond "Seibr-Svitlo" and The Friends of Lviv University, Inc, USA.

Editorial board: Hnatush S., Prokopiv A., Peretyatko T., Halushka A., Bura M., Rabyk M., Yavornytska I., Gnatyna O., Kachmar B., Garayda O., Kotlyarova A., Perejmybida L., Zyn' A., Romanyuk M., Mykytyn N., Pochynok T., Hnatush A., Strus Y.

Scientific committee: prof. Volgin S. O., prof. Gudz S. P., prof Klevets M. Y., prof. Sanagursky D. I., prof. Sybirna N. I., prof. Terek O. I., prof. Fedorenko V. O., prof. Tsaryk Y. V.

   

Institute of Cell Biology of NAS of Ukraine, 14/16, Drahomanov str., Lviv, 79005, Ukraine rosthor@gmail.com In the last decade there has been a significant increase of articles related to giant cell transformation in different cancers. But its still unknown what mechanisms are involved in such giant cancer cells transformation. After vinblastine treatment NK/Ly lymphoma cells undergo giant cell transformation with several cytoskeletal and membrane changes. Our aim was evaluate optimal conditions of NK/Ly-cells permeabilization in order to measure mitochondrial respiration in situ.

NK/Ly lymphoma cell population was cultivated in vivo by intraperitoneal inoculation of 25 million cells into C57 black 4-month mice. Animal weighting was performed every 24 hours. On day 7-10, when the weight increased by 50-60%, mice were drained. To obtain giant cell population (17 mkm), mice were injected 1% vinblastine solution (maximum 50 g) 3 days before drainage.

Cell suspension (approximately 5*106cells/ml) was washed twice from the ascitic fluid with solution containing Cl 90.0 mM, NaCl 15.0 mM, KH2PO4 2 mM, MgCl2 1.0 mM, HEPES10.0 mM, G 0.5 mM, pH 7.2. Afterwards permeabilization with digitonin was performed.

To evaluate concentration of digitonin, sufficient to completely permeabilize plasma membranes of NK/Ly cells, we used it in concentrations 5-150 g/ml/5*106 cells. Cell permeabilization was determined experimentally using trypane blue test and measurement of LDH release using nitrobluetetrasolium. Additionally, we measured respiration rate at 370C of digitonin-permeabilized NK/Ly cells by the polarographic method with Clark oxygen electrode. Respiration was stimulated with ketoglutarate (1 mM), succinate (0.35 mM) and ADP (0.75 mM).

Complete permeabilization of intact NK/Ly cells was achieved after treatment with 20 g/ml of digitonin per 5*106 cells. Surprisingly, giant cells were permeabilized onla after using 150 g/ml digitonin per 5*106 cells. Activity of released LDH after complete permeabilization was three times higher in giant vinblastine treated cells comparing to intact cells. Endogenous respiration rate of intact and giant cells decreased after permeabilization. Substrates and ADP stimulated respiration proportionally to permeabilization degree.

Thus we conclude, that vinblastine treated cells have higher resistance to digitonin. It may by associated with higher cell surface or low cholesterol content, but this needs further investigation.

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