DT-diaphorase and cytochrome B5 reductase in human lung and breast tumours

Marín, A.; Ceráin, Adela López de; Hamilton, Elizabeth; Ad, Lewis; Martinez-Peñuela, JM; Idoate, M.; Bello, J.

doi:10.1038/bjc.1997.485

Cited by 102 publications

(69 citation statements)

References 31 publications

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“…2 NQO1 is a cytosolic enzyme elevated in breast cancers (6) that catalyzes a two-electron reduction of quinones (e.g. ␤-Lap, menadione), utilizing either NADH or NADPH as electron donors.…”

Section: ␤-Lapmentioning

confidence: 99%

“…We propose that development of ␤-Lap for treatment of human cancers that have elevated NQO1 levels (e.g. breast and lung) is warranted (6). Since most clinical agents used to date kill cells by caspase-dependent and p53-dependent pathways, and many cancers evade death by altering these pathways, development of agents that kill by specific targets (NQO1-mediated) and in p53-and caspase-independent manners are needed.…”

Section: Camentioning

confidence: 99%

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Calcium Is a Key Signaling Molecule in β-Lapachone-mediated Cell Death

Tagliarino¹,

Pink²,

Dubyak

et al. 2001

Journal of Biological Chemistry

153

150

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␤-Lapachone (␤-Lap) triggers apoptosis in a number of human breast and prostate cancer cell lines through a unique apoptotic pathway that is dependent upon NQO1, a two-electron reductase. Downstream signaling pathway(s) that initiate apoptosis following treatment with ␤-Lap have not been elucidated. Since calpain activation was suspected in ␤-Lap-mediated apoptosis, we examined alterations in Ca 2؉ homeostasis using NQO1-expressing MCF-7 cells. ␤-Lap-exposed MCF-7 cells exhibited an early increase in intracellular cytosolic Ca 2؉, from endoplasmic reticulum Ca 2؉ stores, comparable to thapsigargin exposures. 1,2-Bis-(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid-acetoxymethyl ester, an intracellular Ca 2؉ chelator, blocked early increases in Ca 2؉ levels and inhibited ␤-Lap-mediated mitochondrial membrane depolarization, intracellular ATP depletion, specific and unique substrate proteolysis, and apoptosis. The extracellular Ca 2؉ chelator, EGTA, inhibited later apoptotic end points (observed >8 h, e.g. substrate proteolysis and DNA fragmentation), suggesting that later execution events were triggered by Ca 2؉ influxes from the extracellular milieu. Collectively, these data suggest a critical, but not sole, role for Ca 2؉ in the NQO1-dependent cell death pathway initiated by ␤-Lap. Use of ␤-Lap to trigger an apparently novel, calpain-like-mediated apoptotic cell death could be useful for breast and prostate cancer therapy. ␤-Lap1 is a naturally occurring compound present in the bark of the South American Lapacho tree. It has antitumor activity against a variety of human cancers, including colon, prostrate, promyelocytic leukemia, and breast (1-3). ␤-Lap was an effective agent (alone and in combination with taxol) against human ovarian and prostate xenografts in mice, with little host toxicity (4). We recently demonstrated that ␤-Lap kills human breast and prostate cancer cells by apoptosis, a cytotoxic response significantly enhanced by NAD(P)H:quinone oxidoreductase (NQO1, E.C. 1.6.99.2) enzymatic activity (5).2 ␤-Lap cytotoxicity was prevented by co-treatment with dicumarol (an NQO1 inhibitor) in NQO1-expressing breast and prostate cancer cells (5).2 NQO1 is a cytosolic enzyme elevated in breast cancers (6) that catalyzes a two-electron reduction of quinones (e.g. ␤-Lap, menadione), utilizing either NADH or NADPH as electron donors. Reduction of ␤-Lap by NQO1 presumably leads to a futile cycling of the compound, wherein the quinone and hydroquinone form a redox cycle with a net concomitant loss of reduced NAD(P)H (5).Apoptosis is an evolutionarily conserved pathway of biochemical and molecular events that underlie cell death processes involving the stimulation of intracellular zymogens. The process is a genetically programmed form of cell death involved in development, normal turnover of cells, and in cytotoxic responses to cellular insults. Once apoptosis is initiated, biochemical and morphological changes occur in the cell. These changes include: DNA fragmentation, chromatin condensation, cytoplasmic membran...

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Section: ␤-Lapmentioning

confidence: 99%

Section: Camentioning

confidence: 99%

Calcium Is a Key Signaling Molecule in β-Lapachone-mediated Cell Death

Tagliarino¹,

Pink²,

Dubyak

et al. 2001

Journal of Biological Chemistry

153

150

View full text Add to dashboard Cite

show abstract

“…Our results, and a previous study (Beall et al, 1995), suggest that this upper threshold may be close to 300 nmol min -1 mg protein -1 . While this may limit the use of this combination therapy approach in some situations, it should not significantly impair the use of DT-diaphorase inducers to increase the anti-tumour activity of bioreductive agents in the clinic since primary tumours generally have base levels of DT-diaphorase that are <100 nmol min -1 mg protein -1 (Schlager and Powis, 1990;Malkinson et al, 1992;Ross et al, 1994;Smitskamp-Wilms et al, 1995;Marin et al, 1997).…”

Section: mentioning

confidence: 99%

Enhanced cytotoxicity of mitomycin C in human tumour cells with inducers of DT-diaphorase

et al. 1999

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Summary DT-diaphorase is a two-electron reducing enzyme that activates the bioreductive anti-tumour agent, mitomycin C (MMC). Cell lines having elevated levels of DT-diaphorase are generally more sensitive to MMC. We have shown that DT-diaphorase can be induced in human tumour cells by a number of compounds, including 1,2-dithiole-3-thione. In this study, we investigated whether induction of DT-diaphorase could enhance the cytotoxic activity of MMC in six human tumour cell lines representing four tumour types. DT-diaphorase was induced by many dietary inducers, including propyl gallate, dimethyl maleate, dimethyl fumarate and sulforaphane. The cytotoxicity of MMC was significantly increased in four tumour lines with the increase ranging from 1.4-to threefold. In contrast, MMC activity was not increased in SK-MEL-28 human melanoma cells and AGS human gastric cancer cells, cell lines that have high base levels of DT-diaphorase activity. Toxicity to normal human marrow cells was increased by 50% when MMC was combined with 1,2-dithiole-3-thione, but this increase was small in comparison with the threefold increase in cytotoxicity to tumour cells. This study demonstrates that induction of DT-diaphorase can increase the cytotoxic activity of MMC in human tumour cell lines, and suggests that it may be possible to use non-toxic inducers of DT-diaphorase to enhance the efficacy of bioreductive anti-tumour agents.

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“…16 DTD is a bioreductive enzyme that can activate a variety of prodrugs, including quinone -based drugs, to DNA alkylating agents. DTD activity has been reported to be elevated in a range of tumor types 30 such as in human non -small cell lung cancer, 13,14 breast, liver, 31 and colon tumors relative to surrounding normal tissue. It has been proposed that various prodrugs that are substrates for DTD could be useful for the treatment of tumors with high DTD activity.…”

Section: Discussionmentioning

confidence: 99%

“…11 The hydroquinone of MMC undergoes chemical rearrangements, resulting in a bifunctional alkylating agent capable of crosslinking DNA. 12 Elevated DTD activities have been reported in tumor relative to normal tissue 13 and in tumor cell lines. 14 It has been suggested that this difference be exploited using compounds that are DTD substrates.…”

mentioning

confidence: 99%

Expression of the prodrug-activating enzyme DT-diaphorase via Ad5 delivery to human colon carcinoma cells in vitro

2002

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Intratumoral injection of recombinant adenoviral type 5 ( Ad5 ) vectors that carry prodrug -activating enzymes like DT -diaphorase ( DTD ) could be used to selectively target tumor cells for chemotherapy. To demonstrate the feasibility of this approach, Ad5 vectors were constructed, which express human DTD minigenes for both wild -type and mutant ( C -to -T change in nucleotide 609 in DTD cDNA ) DTD under the control of the cytomegalovirus ( CMV ) promoter. HT29 human colon carcinoma cells express wild -type DTD, whereas BE human colon carcinoma cells express mutant DTD, have low to undetectable DTD activity, and are 4 -to 6 -fold more resistant to mitomycin C ( MMC ) than HT29 cells. A test of the ability of Ad5 to infect these cells ( using a -galactosidase CMVdriven minigene ) indicated that 90 -100% of BE cells were infected at a multiplicity of infection ( MOI ) of 100, whereas only 15 -40% of HT29 cells were infected at this MOI. Infection of BE cells in vitro with recombinant Ad5 carrying a minigene for wild -type DTD at MOIs of 3 -100 resulted in a progressive increase in DTD activity and a maximal 8 -fold increase in sensitivity to MMC as measured by a colony -forming assay. HT29 cells were sensitized 2 -to 3 -fold following treatment with Ad5.DTD at an MOI of 100. These results indicate that adenovirus -mediated gene transfer and expression of wild -type DTD can sensitize resistant tumor cells to MMC and that this therapeutic strategy may exert a significant bystander effect.

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DT-diaphorase and cytochrome B5 reductase in human lung and breast tumours

Cited by 102 publications

References 31 publications

Calcium Is a Key Signaling Molecule in β-Lapachone-mediated Cell Death

Calcium Is a Key Signaling Molecule in β-Lapachone-mediated Cell Death

Enhanced cytotoxicity of mitomycin C in human tumour cells with inducers of DT-diaphorase

Expression of the prodrug-activating enzyme DT-diaphorase via Ad5 delivery to human colon carcinoma cells in vitro

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