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The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM

Melittin, which acts as a membrane-disrupting lytic peptide, is not only cytotoxic to tumors, but also vital to normal cells. Melittin had low toxicity when coupled with target peptides. Despite significant research development with the fused toxin, a new fused toxin is needed which has a cleavable...

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Autores principales: Sun, Dejun, Sun, Miaonan, Zhu, Wenhe, Wang, Zhiding, Li, Yuefei, Ma, Jie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4344633/
https://www.ncbi.nlm.nih.gov/pubmed/25658509
http://dx.doi.org/10.3390/toxins7020423
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author Sun, Dejun
Sun, Miaonan
Zhu, Wenhe
Wang, Zhiding
Li, Yuefei
Ma, Jie
author_facet Sun, Dejun
Sun, Miaonan
Zhu, Wenhe
Wang, Zhiding
Li, Yuefei
Ma, Jie
author_sort Sun, Dejun
collection PubMed
description Melittin, which acts as a membrane-disrupting lytic peptide, is not only cytotoxic to tumors, but also vital to normal cells. Melittin had low toxicity when coupled with target peptides. Despite significant research development with the fused toxin, a new fused toxin is needed which has a cleavable linker such that the fused toxin can release melittin after protease cleavage on the tumor cell surface. We describe a novel fused toxin, composed of disintegrin, uPA (urokinase-type plasminogen activator)-cleavable linker, and melittin. Disintegrin is a single strand peptide (73 aa) isolated from Gloydius Ussuriensis venom. The RGD (Arg-Gly-Asp) site of disintegrin dominates its interaction with integrins on the surface of the tumor cells. uPA is over-expressed and plays an important role in tumor cell invasiveness and metastatic progression. The DLM (disintegrin-linker-melittin) linker is uPA-cleavable, enabling DLM to release melittin. We compared binding activity of our synthesized disintegrin with native disintegrin and report that DLM had less binding activity than the native form. uPA-cleavage was evaluated in vitro and the uPA-cleavable linker released melittin. Treating tumors expressing uPA with DLM enhanced tumor cell killing as well as reduced toxicity to erythrocytes and other non-cancerous normal cells. The mechanism behind DLM tumor cell killing was tested using a DNA ladder assay, fluorescent microscopy, flow cytometry, and transmission electron microscopy. Data revealed tumor cell necrosis as the mechanism of cell death, and the fused DLM toxin with an uPA-cleavable linker enhanced tumor selectivity and killing ability.
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spelling pubmed-43446332015-03-18 The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM Sun, Dejun Sun, Miaonan Zhu, Wenhe Wang, Zhiding Li, Yuefei Ma, Jie Toxins (Basel) Article Melittin, which acts as a membrane-disrupting lytic peptide, is not only cytotoxic to tumors, but also vital to normal cells. Melittin had low toxicity when coupled with target peptides. Despite significant research development with the fused toxin, a new fused toxin is needed which has a cleavable linker such that the fused toxin can release melittin after protease cleavage on the tumor cell surface. We describe a novel fused toxin, composed of disintegrin, uPA (urokinase-type plasminogen activator)-cleavable linker, and melittin. Disintegrin is a single strand peptide (73 aa) isolated from Gloydius Ussuriensis venom. The RGD (Arg-Gly-Asp) site of disintegrin dominates its interaction with integrins on the surface of the tumor cells. uPA is over-expressed and plays an important role in tumor cell invasiveness and metastatic progression. The DLM (disintegrin-linker-melittin) linker is uPA-cleavable, enabling DLM to release melittin. We compared binding activity of our synthesized disintegrin with native disintegrin and report that DLM had less binding activity than the native form. uPA-cleavage was evaluated in vitro and the uPA-cleavable linker released melittin. Treating tumors expressing uPA with DLM enhanced tumor cell killing as well as reduced toxicity to erythrocytes and other non-cancerous normal cells. The mechanism behind DLM tumor cell killing was tested using a DNA ladder assay, fluorescent microscopy, flow cytometry, and transmission electron microscopy. Data revealed tumor cell necrosis as the mechanism of cell death, and the fused DLM toxin with an uPA-cleavable linker enhanced tumor selectivity and killing ability. MDPI 2015-02-04 /pmc/articles/PMC4344633/ /pubmed/25658509 http://dx.doi.org/10.3390/toxins7020423 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sun, Dejun
Sun, Miaonan
Zhu, Wenhe
Wang, Zhiding
Li, Yuefei
Ma, Jie
The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM
title The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM
title_full The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM
title_fullStr The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM
title_full_unstemmed The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM
title_short The Anti-Cancer Potency and Mechanism of a Novel Tumor-Activated Fused Toxin, DLM
title_sort anti-cancer potency and mechanism of a novel tumor-activated fused toxin, dlm
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4344633/
https://www.ncbi.nlm.nih.gov/pubmed/25658509
http://dx.doi.org/10.3390/toxins7020423
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