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Growth inhibitory activity of a novel lectin from Cliona varians against K562 human erythroleukemia cells

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Abstract

Purpose

In this study, the antitumoral potential of a novel lectin (CvL) purified from the marine sponge Cliona varians was studied in different cancer cell lines.

Methods

CvL cytotoxicity was evaluated in mammalian tumor cells and in normal human peripheral blood lymphocytes by the MTT assay using the same range of concentrations (1–150 µg ml−1). The mechanisms involved in K562 cell death were investigated by confocal fluorescence microscopy, flow cytometry and immunoblot.

Results

CvL inhibited the growth of human leukemia cells, with IC50 values of 70 and 100 µg ml−1 for K562 and JURKAT cells, respectively, but it was ineffective on blood lymphocytes and solid tumor cell lines. K562 cell death occurred 72 h after exposure to the lectin and with signs of apoptosis, as analyzed by DAPI and annexin V/PI staining. Investigation of the possible mediators of this process showed that cell death occurred via a caspase-independent pathway. Confocal fluorescence microscopy indicated a pivotal role for the lysosomal protease cathepsin B in mediating cell death. Accordingly, pre-incubation of K562 cells with the cathepsin inhibitor l-trans-epoxysuccinyl-l-leucylamido-(4-guanidino)butane (E-64) abolished CvL cytotoxic effect. Furthermore, we found upregulation of tumor necrosis factor receptor 1 (TNFR1) and down-modulation of p65 subunit of nuclear factor kappa B (NFκB) expression in CvL-treated cells. These effects were accompanied by increased levels of p21 and reduced expression of pRb, suggesting that CvL can induce cell cycle arrest.

Conclusions

Collectively, these findings indicate an antileukemic effect for CvL and suggest that cathepsin B acts as a death mediator in CvL-induced cytotoxicity possibly in an uncharacterized connection with the membrane death receptor pathway.

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Acknowledgments

The authors acknowledge the support of Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Financiadora de Estudos e Projetos (FINEP) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). The authors thank the Hemonorte, Natal, RN, Brazil, for the generous blood bag donation. R.A.S. was supported by a PhD fellowship from FAPESP (proc. 06/07315-3).

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Authors and Affiliations

  1. Departamento de Biofísica e Farmacologia, Centro de Biociências, Universidade Federal do Rio Grande do Norte (UFRN), CEP 59072-970, Natal, RN, Brazil

    Alexandre F. S. Queiroz & Raniere M. Moura

  2. Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), C.P. 6109, CEP 13083-970, Campinas, SP, Brazil

    Rodrigo A. Silva, Ana C. S. Souza & Giselle Z. Justo

  3. Laboratório de Química e Função de Proteínas Bioativas, Departamento de Bioquímica, Centro de Biociências, UFRN, CEP 59072-970, Natal, RN, Brazil

    Raniere M. Moura & Elizeu A. Santos

  4. Departamento de Bioquímica, Universidade Federal de São Paulo (UNIFESP), Rua 3 de Maio, 100, CEP 04044-020, São Paulo, SP, Brazil

    Juliana L. Dreyfuss, Edgar J. Paredes-Gamero, Ivarne L. S. Tersariol, Helena B. Nader & Giselle Z. Justo

  5. Centro Interdisciplinar de Investigação Bioquímica, Universidade de Mogi das Cruzes, Avenida Dr. Cândido Xavier de Almeida Souza, 200, CEP 08701-970, Mogi das Cruzes, SP, Brazil

    Ivarne L. S. Tersariol

  6. Departamento de Bioquímica, Centro de Biociências, Universidade Federal do Rio Grande do Norte (UFRN), CEP 59072-970, Natal, RN, Brazil

    Maurício P. de Sales

Authors
  1. Alexandre F. S. Queiroz
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  2. Rodrigo A. Silva
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  3. Raniere M. Moura
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  4. Juliana L. Dreyfuss
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  5. Edgar J. Paredes-Gamero
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  6. Ana C. S. Souza
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  7. Ivarne L. S. Tersariol
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  8. Elizeu A. Santos
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  9. Helena B. Nader
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  10. Giselle Z. Justo
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  11. Maurício P. de Sales
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Corresponding authors

Correspondence to Giselle Z. Justo or Maurício P. de Sales.

Additional information

A. F. S. Queiroz and R. A. Silva contributed equally to this manuscript.

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Queiroz, A.F.S., Silva, R.A., Moura, R.M. et al. Growth inhibitory activity of a novel lectin from Cliona varians against K562 human erythroleukemia cells. Cancer Chemother Pharmacol 63, 1023–1033 (2009). https://doi.org/10.1007/s00280-008-0825-4

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  • DOI: https://doi.org/10.1007/s00280-008-0825-4

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