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Загальна кількість знайдених документів : 5
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Gerashchenko O. L. Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells [Електронний ресурс] / O. L. Gerashchenko, E. V. Zhuravel, O. V. Skachkova, N. N. Khranovska, V. V. Filonenko, P. V. Pogrebnoy, M. A. Soldatkina // Experimental oncology. - 2013. - Vol. 35, № 2. - С. 76-82. - Режим доступу: http://nbuv.gov.ua/UJRN/EOL_2013_35_2_1 Aim - the aim of the study was in vitro analysis of biological activity of recombinant human beta-defensin-4 (rec-hBD-4). Methods: hBD-4 cDNA was cloned into pGEX-2T vector, and recombinant plasmid was transformed into E. coli BL21(DE3) cells. To purify soluble fusion GST-hBD-4 protein, affinity chromatography was applied. Rec-hBD-4 was cleaved from the fusion protein with thrombin, and purified by reverse phase chromatography on Sep-Pack C18. Effects of rec-hBD-4 on proliferation, viability, cell cycle distribution, substrate-independent growth, and mobility of cultured human cancer cells of A431, A549, and TPC-1 lines were analyzed by direct cell counting technique, MTT assay, flow cytofluorometry, colony forming assay in semi-soft medium, and wound healing assay. Rech-BD-4 was expressed in bacterial cells as GST-hBD-4 fusion protein, and purified by routine 3-step procedure (affine chromatography on glutathione-agarose, cleavage of fusion protein by thrombin, and reverse phase chromatography). Analysis of in vitro activity of rec-hBD-4 toward three human cancer cell lines has demonstrated that the defensin is capable to affect cell behaviour in concentration-dependent manner. In 1 - 100 nM concentrations rec-hBD-4 significantly stimulates cancer cell proliferation and viability, and promotes cell cycle progression through G2/M checkpoint, greatly enhances colony-forming activity and mobility of the cells. Treatment of the cells with 500 nM of rec-hBD-4 resulted in opposite effects: significant suppression of cell proliferation and viability, blockage of cell cycle in G1/S checkpoint, significant inhibition of cell migration and colony forming activity. Conclusion: recombinant human beta-defensin-4 is biologically active peptide capable to cause oppositely directed effects toward biologic features of cancer cells in vitro dependent on its concentration.
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Gerashchenko O. Involvement of human beta-defensin-2 in regulation of malignant potential of cultured human melanoma cells [Електронний ресурс] / O. Gerashchenko, E. Zhuravel, O. Skachkova, N. Khranovska, V. Pushkarev, P. Pogrebnoy, M. Soldatkina // Experimental oncology. - 2014. - Vol. 36, № 1. - С. 17-23. - Режим доступу: http://nbuv.gov.ua/UJRN/EOL_2014_36_1_5 Background and aim - human beta-defensin-2 (hBD-2) is an antimicrobial cationic peptide capable to control human carcinoma cell growth via cell cycle regulation. The present study was aimed on determination of hBD-2 influence on the growth patterns and malignant potential of cultured human melanoma cells. The study was performed on cultured human melanoma cells of mel Z and mel Is lines treated with recombinant hBD-2 (rec-hBD-2); cell viability, proliferation, cell cycle distribution, and anchorage-independent growth were analyzed using MTT test, direct cell counting, flow cytometry, and colony forming assay respectively. Expression and/or phosphorylation levels of proteins involved in cell cycle control were evaluated by Western blotting. The treatment of mel Z and mel Is cells with rec-hBD-2 in a concentration range of 100 - 1000 nM resulted in a concentration-dependent suppression of cell proliferation, viability, and colony forming activity. It has been shown that rec-hBD-2 exerts its growth suppression effects via significant downregulation of B-Raf expression, activation of pRB and upregulation of p21<^>WAF1 expression, downregulation of cyclin D1 and cyclin E resulting in cell cycle arrest at Gl/S checkpoint. Conclusion: according to obtained results, hBD-2 exerts its growth suppression effect toward human melanoma cells via downregulation of B-Raf, cyclin D1 and cyclin E expression, upregulation of p21<^>WAF1 expression and activation of pRB.
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Gerashchenko O. L. Effect of docetaxel and human beta-defensin-2 on proliferation of аnaplastic thyroid carcinoma KTC-2 cells [Електронний ресурс] / O. L. Gerashchenko, O. V. Zhuravel, V. V. Pushkarev, M. A. Soldatkina, V. M. Pushkarev, P. V. Pogrebnoy // Ендокринологія. - 2014. - Т. 19, № 1. - С. 11-15. - Режим доступу: http://nbuv.gov.ua/UJRN/enkrl_2014_19_1_4 In present research we have studied the effect of docetaxel in uitralow concentrations (0.1-10 nM) and recombinant human beta-defensin-2 (hBD-2) in nanomolar concentration range on proliferation and viability of anaplastic thyroid carcinoma cells of KTC-2 line, and expression of some cell cycle regulatory factors. It has been shown that docetaxel, hBD-2 and combination of these agents significantly inhibited proliferation of KTC-2 cells via down-regulation of the cyclin E expression.Досліджено вплив доцетакселю в наднизьких концентраціях і рекомбінантного бета-дефенсину-2 людини (hBD-2) на проліферативну активність і життєздатність клітин анапластичної карциноми щитовидної залози лінії КТС-2 та на експресію деяких регуляторних факторів клітинного циклу. Показано, що дія доцетакселю, hBD-2 та комбінації цих двох агентів призводила до значного зниження проліферації клітин КТС-2 шляхом пригнічення експресії цикліну Е.В работе исследовано влияние доцетакселя в сверхнизких концентрациях (0,1-10 нМ) и рекомбинантного бета-дефенсина-2 человека (hBD-2) в наномолярном диапазоне концентраций на пролиферативную активность и жизнеспособность клеток анапластической карциномы щитовидной железы линииКТС-2 и экспрессию некоторых регуляторных факторов клеточного цикла. Показано, что действие доцетакселя, hBD-2 и их комбинации приводит к значительному угнетению пролиферации клеток КТС-2 путем снижения экспрессии циклина Е.
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Zhuravel O. V. Expression of human beta-defensins-1–4 in thyroid cancer cells and new insight on biologic activity of hBD-2 in vitro [Електронний ресурс] / O. V. Zhuravel, O. L. Gerashchenko, M. R. Khetsuriani, M. A. Soldatkina, P. V. Pogrebnoy // Experimental oncology. - 2014. - Vol. 36, № 3. - С. 174-178. - Режим доступу: http://nbuv.gov.ua/UJRN/EOL_2014_36_3_7
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Gerashchenko O. Fusion expression of recombinant human beta-defensin-3 and analysis of its biological activity [Електронний ресурс] / O. Gerashchenko, V. Kovalchuk, O. Boidunik, M. Soldatkina, P. Pogrebnoy // Вісник Київського національного університету імені Тараса Шевченка. Біологія. - 2014. - Вип. 3. - С. 98-102. - Режим доступу: http://nbuv.gov.ua/UJRN/VKNU_biol_2014_3_28
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