NAD(P)H: quinone oxidoreductase activity is the principal determinant of β-lapachone cytotoxicity

John J. Pink, Sarah M. Planchon, Colleen Tagliarino, Marie E. Varnes, David Siegel, David A. Boothman

Research output: Contribution to journalArticle

270 Citations (Scopus)

Abstract

β-Lapachone activates a novel apoptotic response in a number of cell lines. We demonstrate that the enzyme NAD(P)H:quinone oxidoreductase (NQO1) substantially enhances the toxicity of β-lapachone. NQO1 expression directly correlated with sensitivity to a 4-h pulse of β-lapachone in a panel of breast cancer cell lines, and the NQO1 inhibitor, dicoumarol, significantly protected NQO1-expressing cells from all aspects of β-lapachone toxicity. Stable transfection of the NQO1-deficient cell line, MDA-MB-468, with an NQO1 expression plasmid increased apoptotic responses and lethality after β- lapachone exposure. Dicoumarol blocked both the apoptotic responses and lethality. Biochemical studies suggest that reduction of β-lapachone by NQO1 leads to a futile cycling between the quinone and hydroquinone forms, with a concomitant loss of reduced NAD(P)H. In addition, the activation of a cysteine protease, which has characteristics consistent with the neutral calcium-dependent protease, calpain, is observed after β-lapachone treatment. This is the first definitive elucidation of an intracellular target for β-lapachone in tumor cells. NQO1 could be exploited for gene therapy, radiotherapy, and/or chemopreventive interventions, since the enzyme is elevated in a number of tumor types (i.e. breast and lung) and during neoplastic transformation.

Original languageEnglish (US)
Pages (from-to)5416-5424
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number8
DOIs
StatePublished - Feb 25 2000

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Cytotoxicity
NAD
Dicumarol
Oxidoreductases
Calpain
Cells
Cell Line
Substrate Cycling
Breast Neoplasms
Toxicity
Tumors
Cysteine Proteases
Enzymes
Gene therapy
Genetic Therapy
Transfection
Radiotherapy
Plasmids
Lung
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pink, J. J., Planchon, S. M., Tagliarino, C., Varnes, M. E., Siegel, D., & Boothman, D. A. (2000). NAD(P)H: quinone oxidoreductase activity is the principal determinant of β-lapachone cytotoxicity. Journal of Biological Chemistry, 275(8), 5416-5424. https://doi.org/10.1074/jbc.275.8.5416

NAD(P)H : quinone oxidoreductase activity is the principal determinant of β-lapachone cytotoxicity. / Pink, John J.; Planchon, Sarah M.; Tagliarino, Colleen; Varnes, Marie E.; Siegel, David; Boothman, David A.

In: Journal of Biological Chemistry, Vol. 275, No. 8, 25.02.2000, p. 5416-5424.

Research output: Contribution to journalArticle

Pink, JJ, Planchon, SM, Tagliarino, C, Varnes, ME, Siegel, D & Boothman, DA 2000, 'NAD(P)H: quinone oxidoreductase activity is the principal determinant of β-lapachone cytotoxicity', Journal of Biological Chemistry, vol. 275, no. 8, pp. 5416-5424. https://doi.org/10.1074/jbc.275.8.5416
Pink, John J. ; Planchon, Sarah M. ; Tagliarino, Colleen ; Varnes, Marie E. ; Siegel, David ; Boothman, David A. / NAD(P)H : quinone oxidoreductase activity is the principal determinant of β-lapachone cytotoxicity. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 8. pp. 5416-5424.
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