Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells
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 chr...
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| Published in: | Experimental Oncology |
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| Date: | 2013 |
| Main Authors: | , , , , , , |
| Format: | Article |
| Language: | English |
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2013
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| Online Access: | https://nasplib.isofts.kiev.ua/handle/123456789/145208 |
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| Journal Title: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Cite this: | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells / O.L. Gerashchenko, E.V. Zhuravel, O.V. Skachkova, N.N. Khranovska, V.V. Filonenko, P.V. Pogrebnoy, A. Soldatkina // Experimental Oncology. — 2013. — Т. 35, № 2. — С. 76–82. — Бібліогр.: 37 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1862646027347034112 |
|---|---|
| author | Gerashchenko, O.L. Zhuravel, E.V. Skachkova, O.V. Khranovska, N.N. Filonenko, V.V. Pogrebnoy, P.V. Soldatkina, M.A. |
| author_facet | Gerashchenko, O.L. Zhuravel, E.V. Skachkova, O.V. Khranovska, N.N. Filonenko, V.V. Pogrebnoy, P.V. Soldatkina, M.A. |
| citation_txt | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells / O.L. Gerashchenko, E.V. Zhuravel, O.V. Skachkova, N.N. Khranovska, V.V. Filonenko, P.V. Pogrebnoy, A. Soldatkina // Experimental Oncology. — 2013. — Т. 35, № 2. — С. 76–82. — Бібліогр.: 37 назв. — англ. |
| collection | DSpace DC |
| container_title | Experimental Oncology |
| description | 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.
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| first_indexed | 2025-12-01T10:51:05Z |
| format | Article |
| fulltext | |
| id | nasplib_isofts_kiev_ua-123456789-145208 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-12-01T10:51:05Z |
| publishDate | 2013 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Gerashchenko, O.L. Zhuravel, E.V. Skachkova, O.V. Khranovska, N.N. Filonenko, V.V. Pogrebnoy, P.V. Soldatkina, M.A. 2019-01-19T11:59:47Z 2019-01-19T11:59:47Z 2013 Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells / O.L. Gerashchenko, E.V. Zhuravel, O.V. Skachkova, N.N. Khranovska, V.V. Filonenko, P.V. Pogrebnoy, A. Soldatkina // Experimental Oncology. — 2013. — Т. 35, № 2. — С. 76–82. — Бібліогр.: 37 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/145208 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. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells Article published earlier |
| spellingShingle | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells Gerashchenko, O.L. Zhuravel, E.V. Skachkova, O.V. Khranovska, N.N. Filonenko, V.V. Pogrebnoy, P.V. Soldatkina, M.A. Original contributions |
| title | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells |
| title_full | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells |
| title_fullStr | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells |
| title_full_unstemmed | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells |
| title_short | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells |
| title_sort | biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells |
| topic | Original contributions |
| topic_facet | Original contributions |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/145208 |
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