Laccase purified from Cerrena unicolor exerts antitumor activity against leukemic cells

Matuszewska,Anna; Karp,Marta; Jaszek,Magdalena; Janusz,Grzegorz; Osińska‑Jaroszuk,Monika; Sulej,Justyna; Stefaniuk,Dawid; Tomczak,Waldemar; Giannopoulos,Krzysztof

doi:10.3892/ol.2016.4220

Laccase purified from Cerrena unicolor exerts antitumor activity against leukemic cells

Authors:
- Anna Matuszewska
- Marta Karp
- Magdalena Jaszek
- Grzegorz Janusz
- Monika Osińska‑Jaroszuk
- Justyna Sulej
- Dawid Stefaniuk
- Waldemar Tomczak
- Krzysztof Giannopoulos
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Affiliations: Department of Biochemistry, Maria Curie‑Skłodowska University, Lublin 20‑033, Poland, Department of Experimental Hemato‑Oncology, Medical University of Lublin, Lublin 20‑093, Poland, Department of Hemato‑Oncology and Bone Marrow Transplantation, Medical University of Lublin, Lublin 20‑081, Poland
Published online on: February 9, 2016 https://doi.org/10.3892/ol.2016.4220
Pages: 2009-2018
Copyright: © Matuszewska et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Chronic lymphocytic leukemia (CLL) is the most commonly observed adult hematological malignancy in Western countries. Despite the fact that recent improvements in CLL treatment have led to an increased percentage of complete remissions, CLL remains an incurable disease. Cerrena unicolor is a novel fungal source of highly active extracellular laccase (ex‑LAC) that is currently used in industry. However, to the best of our knowledge, no reports regarding its anti‑leukemic activity have been published thus far. In the present study, it was hypothesized that C. unicolor ex‑LAC may possess cytotoxic activity against leukemic cell lines and CLL primary cells. C. unicolor ex‑LAC was separated using anion exchange chromatography on diethylaminoethyl cellulose‑Sepharose and Sephadex G‑50 columns. The cytotoxic effects of ex‑LAC upon 24‑ and 48‑h treatment on HL‑60, Jurkat, RPMI 8226 and K562 cell lines, as well as CLL primary cells of nine patients with CLL, were evaluated using 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay. Annexin V/propidium iodide staining of Jurkat cells treated with ex‑LAC was used to investigate apoptosis via flow cytometry. Ex‑LAC induced changes in Jurkat and RPMI 8226 cells, as visualized by fluorescence and scanning electron microscopy (SEM). The XTT assay revealed high cytotoxic rates following treatment with various concentrations of ex‑LAC on all the cell lines and CLL primary cells analyzed, with a half maximal inhibitory concentration ranging from 0.4 to 1.1 µg/ml. Fluorescence microscopy and SEM observations additionally revealed apoptotic changes in Jurkat and RPMI 8226 cells treated with ex‑LAC, compared with control cells. These results were in agreement with the apoptosis analysis of Jurkat cells on flow cytometry. In conclusion, C. unicolor ex‑LAC was able to significantly induce cell apoptosis, and may represent a novel therapeutic agent for the treatment of various hematological neoplasms.

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