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 Blood, 1 September 2004, Vol. 104, No. 5, pp. 1459-1464.
Prepublished online as a Blood First Edition Paper on May 6, 2004; DOI 10.1182/blood-2004-02-0594.

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NEOPLASIA

Molecular mechanism of violacein-mediated human leukemia cell death

Carmen Veríssima Ferreira, Carina L. Bos, Henri H. Versteeg, Giselle Z. Justo, Nelson Durán, and Maikel P. Peppelenbosch

From the Departamento de Bioquímica, Universidade Estadual de Campinas, São Paulo, Brazil; the Department of Experimental Internal Medicine, Academic Medical Center, Amsterdam, The Netherlands; the Biological Chemistry Laboratory, Instituto de Química, Universidade Estadual de Campinas, São Paulo, Brazil; and the Núcleo de Ciências Ambientais, Universidade de Mogi das Cruzes, São Paulo, Brazil.

Violacein, a pigment isolated from Chromobacterium violaceum in the Amazon River, presents diverse biologic properties and attracts interest as a consequence of its antileukemic activity. Elucidation of the molecular mechanism mediating this activity will provide further relevant information for understanding its effects on the cellular physiology of untransformed cells and for considering its possible clinical application. Here, we show that violacein causes apoptosis in HL60 leukemic cells but is ineffective in this respect in other types of leukemia cells or in normal human lymphocytes and monocytes. Violacein cytotoxicity in HL60 cells was preceded by activation of caspase 8, transcription of nuclear factor {kappa}B (NF-{kappa}B) target genes, and p38 mitogen-activated protein (MAP) kinase activation. Thus, violacein effects resemble tumor necrosis factor {alpha} (TNF-{alpha}) signal transduction in these cells. Accordingly, infliximab, an antibody that antagonizes TNF-{alpha}–induced signaling abolished the biologic activity of violacein. Moreover, violacein directly activated TNF receptor 1 signaling, because a violacein-dependent association of TNF receptor-associated factor 2 (TRAF2) to this TNF receptor was observed in coimmunoprecipitation experiments. Hence, violacein represents the first member of a novel class of cytotoxic drugs mediating apoptosis of HL60 cells by way of the specific activation of TNF receptor 1.


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