Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells.
| Title: | Biologic activities of recombinant human-beta-defensin-4 toward cultured human cancer cells. |
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| Authors: | Gerashchenko OL; R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv 03022, Ukraine.; Zhuravel EV; Skachkova OV; Khranovska NN; Filonenko VV; Pogrebnoy PV; Soldatkina MA |
| Source: | Experimental oncology [Exp Oncol] 2013 Jun; Vol. 35 (2), pp. 76-82. |
| Publication Type: | Journal Article; Research Support, Non-U.S. Gov't |
| Language: | English |
| Journal Info: | Publisher: Akademperiodyka Country of Publication: Ukraine NLM ID: 101230541 Publication Model: Print Cited Medium: Internet ISSN: 2312-8852 (Electronic) Linking ISSN: 18129269 NLM ISO Abbreviation: Exp Oncol Subsets: MEDLINE |
| Imprint Name(s): | Publication: [Kyiv] : Akademperiodyka; Original Publication: Kyiv : Morion, [2004]- |
| MeSH Terms: | Antineoplastic Agents/*pharmacology ; beta-Defensins/*genetics ; beta-Defensins/*pharmacology; Cell Line, Tumor/drug effects ; Cell Movement/drug effects ; Cell Movement/genetics ; Cell Proliferation/drug effects ; Cell Survival/drug effects ; Protein Engineering/methods ; Recombinant Fusion Proteins/pharmacology ; Dose-Response Relationship, Drug ; Humans ; Tumor Stem Cell Assay |
| Abstract: | 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.; Results: Rec-hBD-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. |
| Substance Nomenclature: | 0 (Antineoplastic Agents); 0 (DEFB4A protein, human); 0 (Recombinant Fusion Proteins); 0 (beta-Defensins) |
| Entry Date(s): | Date Created: 20130706 Date Completed: 20140127 Latest Revision: 20130705 |
| Update Code: | 20260130 |
| PMID: | 23828379 |
| Database: | MEDLINE |
Journal Article; Research Support, Non-U.S. Gov't