Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells

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...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Experimental Oncology
Datum:2013
Hauptverfasser: Gerashchenko, O.L., Zhuravel, E.V., Skachkova, O.V., Khranovska, N.N., Filonenko, V.V., Pogrebnoy, P.V., Soldatkina, M.A.
Format: Artikel
Sprache:English
Veröffentlicht: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2013
Schlagworte:
Original contributions
Online Zugang:https://nasplib.isofts.kiev.ua/handle/123456789/145208
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
Назва журналу:Digital Library of Periodicals of National Academy of Sciences of Ukraine
Zitieren: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.

Institution

Digital Library of Periodicals of National Academy of Sciences of Ukraine
id nasplib_isofts_kiev_ua-123456789-145208
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
institution Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection DSpace DC
title Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells
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_short 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_sort biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells
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.
topic Original contributions
topic_facet Original contributions
publishDate 2013
language English
container_title Experimental Oncology
publisher Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
format Article
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.
issn 1812-9269
url https://nasplib.isofts.kiev.ua/handle/123456789/145208
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.
work_keys_str_mv AT gerashchenkool biologicactivitiesofrecombinanthumanbetadefensin4towardculturedhumancancercells
AT zhuravelev biologicactivitiesofrecombinanthumanbetadefensin4towardculturedhumancancercells
AT skachkovaov biologicactivitiesofrecombinanthumanbetadefensin4towardculturedhumancancercells
AT khranovskann biologicactivitiesofrecombinanthumanbetadefensin4towardculturedhumancancercells
AT filonenkovv biologicactivitiesofrecombinanthumanbetadefensin4towardculturedhumancancercells
AT pogrebnoypv biologicactivitiesofrecombinanthumanbetadefensin4towardculturedhumancancercells
AT soldatkinama biologicactivitiesofrecombinanthumanbetadefensin4towardculturedhumancancercells
first_indexed 2025-12-01T10:51:05Z
last_indexed 2025-12-01T10:51:05Z
_version_ 1850860036911267840

Ähnliche Einträge