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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References
Adams, G. E. & Stratford, I. J. (1994). Bioreductive drugs for cancer therapy: the search for tumor specificity. Int J Radiat Oncol Biol Phys 29: 231–238.
Appelt, L. C. & Reicks, M. M. (1997). Soy feeding induces phase II enzymes in rat tissues. Nutr Cancer 28: 270–275.
Barham, H. M., Inglis, R., Chinje, E. C. & Stratford, I. J. (1996). Development and validation of a spectrophotometric assay for measuring the activity of NADH:cytochrome b5 reductase in human tumour cells. Br J Cancer 74: 1188–1193.
Beall, H. D., Murphey, A. M., Siegel, D., Hargreaves, R. H., Butler, J. & Ross, D. (1995). Nicotinamide adenine dinucleotide (phosphate): quinone oxidoreductase (DT-diaphorase) as a target for bioreductive antitumour quinones: quinone cytotoxicity and selectivity in human lung and breast cancer cell lines. Mol Pharmacol 48: 499–504.
Beall, H. D., Liu, Y. F., Siegel, D., Bolton, E. M., Gibson, N. W. & Ross, D. (1996). Role of NAD(P)H:quinone oxidoreductase (DT-diaphorase) in cytotoxicity and induction of DNA damage by streptonigrin. Biochem Pharmacol 51: 645–652.
Bedikian, A. Y., Legha, S. S., Eton, O., Buzaid, A. C., Papadopoulos, N., Coates, S., Simmons, T., Neefe, J. & von Roemeling, R. (1997). Phase II trial of tirapazamine combined with cisplatin in chemotherapy of advanced malignant melanoma. Ann Oncol 8: 363–367.
Begleiter, A., Robotham, E., Lacey, G. & Leith, M. K. (1989). Increased sensitivity of quinone resistant cells to mitomycin C. Cancer Lett 45: 173–176.
Begleiter, A., Robotham, E. & Leith, M. K. (1992). The role of NAD(P)H:(quinone acceptor) oxidoreductase (DT-diaphorase) in activation of mitomycin C under hypoxia. Mol Pharmacol 41: 677–683.
Begleiter, A., Verburg, L., Ashique, A., Lee, K., Israels, L. G., Mowat, M. R. A. & Johnston, J. B. (1995). Comparison of antitumor activities of 2-chlorodeoxyadenosine and 9-β-arabinosyl-2-fluoroadenine in chronic lymphocytic leukemia and marrow cells in vitro. Leukemia 9: 1875–1881.
Begleiter, A., Leith, M. K. & Curphey, T. J. (1996). Induction of DT-diaphorase by 1,2-dithiole-3-thione and increase of antitumour activity of bioreductive agents. Br J Cancer 74: S9–S14.
Belcourt, M. F., Hodnick, W. F., Rockwell, S. & Sartorelli, A. C. (1996). Differential toxicity of mitomycin C and porfiromycin to aerobic and hypoxic Chinese hamster ovary cells overexpressing human NADPH:cytochrome C (P450) reductase. Proc Natl Acad Sci USA 93: 456–460.
Belinsky, M. & Jaiswal, A. L. (1993). NAD(P)H:quinone oxidoreductase 1 (DT-diaphorase) expression in normal and tumor tissues. Cancer Metastasis Rev 12: 103–117.
Beyer, R. E., Segura Aguilar, J. E. & Ernster, L. (1988). The anticancer enzyme DT diaphorase is induced selectively in liver during ascites hepatoma growth. Anticancer Res 8: 233–238.
Boyer, M. J. (1997). Bioreductive agents: a clinical update. Oncol Res 9: 391–395.
Coia, L. R. (1993). The use of mitomycin in esophageal cancer. Oncology 50: 53–62.
Cummings, B. J., Keane, T. J., O’Sullivan, B., Wong, C. S. & Catton, C. N. (1993). Mitomycin in anal canal carcinoma. Oncology 50: 63–69.
Doherty, G. P., Leith, M. K., Wang, X., Curphey, T. J. & Begleiter, A. (1998). Induction of DT-diaphorase by 1,2-dithiole-3-thiones in human tumour and normal cells and effect on anti-tumour activity of bioreductive agents. Br J Cancer 77: 1241–1252.
Egner, P. A., Kensler, T. W., Prestera, T., Talalay, P., Libby, A. H., Joyner, H. H. & Curphey, T. J. (1994). Regulation of phase 2 enzyme induction by oltipraz and other dithiolethiones. Carcinogenesis 15: 177–181.
Fitzsimmons, S. A., Workman, P., Grever, M., Paull, K., Camalier, R. & Lewis, A. D. (1996). Reductase enzyme expression across the National Cancer Institute tumor cell line panel: correlation with sensitivity to mitomycin C and EO9. J Natl Cancer Inst 88: 259–269.
Fujita, K., Kubota, T., Matsuzaki, S. W., Otani, Y., Watanabe, M., Teramoto, T., Kumai, K. & Kitajima, M. (1998). Further evidence for the value of the chemosensitivity test in deciding appropriate chemotherapy for advanced gastric cancer. Anticancer Res 18: 1973–1978.
Gustafson, D. L. & Pritsos, C. A. (1992). Bioactivation of mitomycin C by xanthine dehydrogenase from EMT6 mouse mammary carcinoma tumors. J Natl Cancer Inst 84: 1180–1185.
Habig, W. H., Pabst, M. J. & Jakoby, W. B. (1974). Glutathione S-transferase. The first enzymatic step in mercapturic acid formation. J Biol Chem 249: 7130–7139.
Hodnick, W. F. & Sartorelli, A. C. (1993). Reductive activation of mitomycin C by NADH:cytochrome b5 reductase. Cancer Res 53: 4907–4912.
Hortobagyi, G. N. (1993). Mitomycin: its evolving role in the treatment of breast cancer. Oncology 50: 1–8.
Jaiswal, A. K. (1991). Human NAD(P)H:quinone oxidoreductase (NQO-1) gene structure and induction by dioxin. Biochemistry 30: 10647–10653.
Jaiswal, A. K., Burnett, P., Adesnik, M. & McBride, O. W. (1990). Nucleotide and deduced amino acid sequence of a human cDNA (NQO2) corresponding to a second member of the NAD(P)H:quinone oxidoreductase gene family. Extensive polymorphism at the NQO2 gene locus on chromosome 6. Biochemistry 29: 1899–1906.
Johnston, J. B., Verburg, L., Shore, T., Williams, M., Israels, L. G. & Begleiter, A. (1994). Combination therapy with nucleoside analogs and alkylating agents. Leukemia 8: S140–S143.
Kensler, T. W. & Helzlsouer, K. J. (1995). Oltipraz: clinical opportunities for cancer chemoprevention. J Cell Biochem Suppl. 22: 101–107.
Kirkpatrick, D. L., Duke, M. & Goh, T. S. (1990). Chemosensitivity testing of fresh human leukemia cells using both a dye exclusion assay and a tetrazolium dye (MTT) assay. Leuk Res 14: 459–466.
Lown, J. W., Begleiter, A., Johnson, D. & Morgan, A. R. (1976). Studies related to antitumor antibiotics. Part V. Reaction of mitomycin C with DNA examined by ethidium fluorescence assay. Can J Biochem 54: 110–119.
Malkinson, A. M., Siegel, D., Forrest, G. L., Gazdar, A. F., Oie, H. K., Chan, D. C., Bunn, P. A., Mabry, M., Dykes, D. J., Harrison, S. D. & Ross, D. (1992). Elevated DT-diaphorase activity and messenger RNA content in human non-small cell lung carcinoma: relationship to the response of lung tumor xenografts to mitomycin C. Cancer Res 52: 4752–4757.
Manson, M. M., Ball, H. W., Barrett, M. C., Clark, H. L., Judah, D. J., Williamson, G. & Neal, G. E. (1997). Mechanism of action of dietary chemoprotective agents in rat liver: induction of phase I and II drug metabolizing enzymes and aflatoxin B1 metabolism. Carcinogenesis 18: 1729–1738.
Marin, A., Lopez de Cerain, A., Hamilton, E., Lewis, A. D., Martinez-Penuela, J. M., Idoate, M. A. & Bello, J. (1997). DT-diaphorase and cytochrome b5 reductase in human lung and breast tumours. Br J Cancer 76: 923–929.
Marshall, R. S. & Rauth, A. M. (1986). Modification of the cytotoxic activity of mitomycin C by oxygen and ascorbic acid in Chinese hamster ovary cells and a repair-deficient mutant. Cancer Res 46: 2709–2713.
Mikami, K., Naito, M., Tomida, A., Yamada, M., Sirakusa, T. & Tsuruo, T. (1996). DT-diaphorase as a critical determinant of sensitivity to mitomycin C in human colon and gastric carcinoma cell lines. Cancer Res 56: 2823–2826.
Miller, V. A., Ng, K. K., Grant, S. C., Kindler, H., Pizzo, B., Heelan, R. T., von Roemeling, R. & Kris, M. G. (1997). Phase II study of the combination of the novel bioreductive agent, tirapazamine, with cisplatin in patients with advanced non small cell lung cancer. Ann Oncol 8: 1269–1271.
Nishiyama, M., Saeki, S., Aogi, K., Hirabayashi, N. & Toge, T. (1993). Relevance of DT-diaphorase activity to mitomycin C (MMC) efficacy on human cancer cells: differences in in vitro and in vivo systems. Int J Cancer 53: 1013–1016.
Nishiyama, M., Suzuki, K., Kumazaki, T., Yamamoto, W., Toge, T., Okamura, T. & Kurisu, K. (1997). Molecular targeting of mitomycin C chemotherapy. Int J Cancer 72: 649–656.
Plumb, J. A., Gerritsen, M., Milroy, R., Thomson, P. & Workman, P. (1994). Relative importance of DT-diaphorase and hypoxia in the bioactivation of EO9 by human lung tumor cell lines. Int J Radiat Oncol Biol Phys 29: 295–299.
Prestera, T., Holtzclaw, W. D., Zhang, Y. & Talalay, P. (1993). Chemical and molecular regulation of enzymes that detoxify carcinogens. Proc Natl Acad Sci USA 90: 2965–2969.
Pritsos, C. A. & Sartorelli, A. C. (1986). Generation of reactive oxygen radicals through bioactivation of mitomycin C antibiotics. Cancer Res 46: 3528–3532.
Prochaska, H. J. & Santamaria, A. B. (1988). Direct measurement of NAD(P)H:quinone reductase from cells cultured in microtiter wells: a screening assay for anticarcinogenic enzyme inducers. Anal Biochem 169: 328–336.
Riley, R. J. & Workman, P. (1992). DT-diaphorase and cancer chemotherapy. Biochem Pharmacol 43: 1657–1669.
Rockwell, S., Sartorelli, A. C., Tomasz, M. & Kennedy, K. A. (1993). Cellular pharmacology of quinone bioreductive alkylating agents. Cancer Metastasis Rev 12: 165–176.
Ross, D., Siegel, D., Beall, H., Prakash, A. S., Mulcahy, R. T. & Gibson, N. W. (1993). DT-diaphorase in activation and detoxification of quinones. Cancer Metastasis Rev 12: 83–101.
Ross, D., Beall, H., Traver, R. D., Siegel, D., Phillips, R. M. & Gibson, N. W. (1994). Bioactivation of quinones by DT-diaphorase, molecular, biochemical, and chemical studies. Oncol Res 6: 493–500.
Schellens, J. H. M., Planting, A. S. T., Van Acker, B. A. C., Loos, W. J., De Boer-Dennert, M., Van der Burg, M. E. L., Koier, I., Krediet, R. T., Stoter, G. & Verweij, J. (1994). Phase I and pharmacologic study of the novel indoloquinone bioreductive alkylating cytotoxic drug EO9. J Natl Cancer Inst 86: 906–912.
Schlager, J. J. & Powis, G. (1990). Cytosolic NAD(P)H: (quinone acceptor) oxidoreductase in human normal and tumor tissue: effects of cigarette smoking and alcohol. Int J Cancer 45: 403–409.
Smitskamp-Wilms, E., Giaccone, G., Pinedo, H. M., Van der Laan, BFAM & Peters, G. J. (1995). DT-diaphorase activity in normal and neoplastic human tissues: an indicator of sensitivity to bioreductive agents?. Br J Cancer 72: 917–921.
Spain, R. C. (1993). The case for mitomycin in non-small cell lung cancer. Oncology 50: 35–52.
Strobel, H. W. & Dignam, J. D. (1978). Purification and properties of NADPH-cytochrome P-450 reductase. Methods Enzymol 52: 89–96.
Tomasz, M., Lipman, R., Chowdary, D., Pawlak, L., Verdine, G. L. & Nakanishi, K. (1987). Isolation and structure of a covalent cross-link adduct between mitomycin C and DNA. Science 235: 1204–1208.
Wang, W. & Higuchi, C. M. (1995). Induction of NAD(P)H:quinone reductase by vitamins A, E, and C in Colo205 cancer cells. Cancer Lett 98: 63–69.
Winski, S. L., Hargreaves, R. H. J., Butler, J. & Ross, D. (1998). New aziridinylbenzoquinones as NAD(P)H:quinone oxidoreductase (NQO1)-directed antitumor agents. Proc Am Assoc Cancer Res 29: 303
Workman, P. & Stratford, I. J. (1993). The experimental development of bioreductive drugs and their role in cancer therapy. Cancer Metastasis Rev 12: 73–82.
Xu, B. H., Gupta, V. & Singh, S. V. (1994). Characterization of a human bladder cancer cell line selected for resistance to mitomycin C. Int J Cancer 58: 686–692.
Zhang, Y., Kensler, T. W., Cho, C. G., Posner, G. H. & Talalay, P. (1994). Anticarcinogenic activities of sulforaphane and structurally related synthetic norbornyl isothiocyanates. Proc Natl Acad Sci USA 91: 3147–3150.
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Wang, X., Doherty, G., Leith, M. et al. Enhanced cytotoxicity of mitomycin C in human tumour cells with inducers of DT-diaphorase. Br J Cancer 80, 1223–1230 (1999). https://doi.org/10.1038/sj.bjc.6690489
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DOI: https://doi.org/10.1038/sj.bjc.6690489
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