© 1996 by Oxford University Press
Journal of the National Cancer Institute, Vol. 88, No. 5, 259-269,
March 6, 1996
© 1996 Oxford University Press
Reductase Enzyme Expression Across the National Cancer Institute Tumor Cell Line Panel: Correlation With Sensitivity to Mitomycin C and EO9
Cancer Research Campaign (CRC) Beatson Laboratories, CRC Department of Medical Oncology, University of Glasgow Scotland, U.K.
Information Technology Branch, Division of Cancer Treatment. National Cancer Institute Bethesda, MD
Correspondence to present address: Paul Workman, Ph.D., ZENECA Pharmaceuticals, Cancer Research Department. Mereside. Alderley Park, Macclesfield, Cheshire, U.K. SK 10 4JG.
BACKGROUND: Many antitumor drugs require metabolic activation to exert their cytotoxic or cytostatic effects. The socalled bioreductive compounds, whose conversion into active antitumor agents is catalyzed by reductase enzymes, are examples of such drugs. The identification of specific enzymes involved in the activation of these compounds is important in understanding cellular factors that may influence drug antitumor activity.
PURPOSE: We measured expression levels of three different reductase enzymes—DT-diaphorase [NAD(P)H (i.e., reduced nicotinamide adenine dinucleotide, with or without phosphate): quinone oxidoreductase]; NADPH: cytochrome P-450 reductase; and NADH (i.e., reduced nicotinamide adenine dinucleotide): cytochrome-b5 reductase—in 69 cell lines (most of the National Cancer Institute [NCI] human tumor cell panel) to see if relationships could be established between the activities of these enzymes and cellular sensitivities to the bioreductive compounds mitomycin C and EO9.
METHODS: For all 69 cell lines, the activity of each enzyme was determined using cellular extracts and photometric assays involving the reduction of cytochrome c. Western blot analysis was used to measure the relative amount of DT-diaphorase protein in each extract, and coupled reverse transcription and polymerase chain reactions were employed to assess DT-diaphorase and NADPH: cytochrome P-450 reductase messenger RNA (mRNA) levels in a subset of the cell lines. The cytotoxic and/or cytostatic activities of mitomycin C and EO9 toward the cell lines were determined under aerobic conditions. Relationships between enzyme activity levels and drug sensitivities were assessed by use of the COMPARE program and Pearson correlation coefficients.
RESULTS: In general, DT-diaphorase activity levels were higher than those observed for the other two reductases across the entire cell line panel. Measured activities for all three enzymes varied among cell lines derived from the same tissue as well as between lines derived from different tissues; however, tissuespecific patterns of expression could be discerned. Differences in the activity levels of individual enzymes appeared to reflect differences in corresponding enzyme protein and/or mRNA levels. A relationship between enzyme activity and chemosensitivities to mitomycin C and EO9 was observed only for DT-diaphorase (Pearson correlation coefficient =.424 [two-sided P<.0005] for mitomycin C and.446 [two-sided P
.0013] for EO9).
CONCLUSIONS: Reductase enzyme expression is heterogeneous across human tumor cell lines, and tissue-specific patterns of expression are apparent. DT-diaphorase activity levels correlate with sensitivities to mitomycin C and EO9, supporting a role for this enzyme in the bioactivation of these anticancer compounds.
IMPLICATIONS: Comparison of biochemical, molecular biological, and chemosensitivity data obtained from screening a large number of cell lines (e.g., the NCI tumor cell line panel) may facilitate investigation of factors influencing drug antitumor activity. The knowledge gained may be of value in the development of new anticancer agents or in the selection of patients to receive specific therapies. [J Natl Cancer Inst 1996;88:259–69]
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  C. Yan, J. K. Kepa, D. Siegel, I. J. Stratford, and D. Ross Dissecting the Role of Multiple Reductases in Bioactivation and Cytotoxicity of the Antitumor Agent 2,5-Diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone (RH1) Mol. Pharmacol., December 1, 2008; 74(6): 1657 - 1665. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Jamieson, K. Wilson, S. Pridgeon, J. P. Margetts, R. J. Edmondson, H. Y. Leung, R. Knox, and A. V. Boddy NAD(P)H:Quinone Oxidoreductase 1 and NRH:Quinone Oxidoreductase 2 Activity and Expression in Bladder and Ovarian Cancer and Lower NRH:Quinone Oxidoreductase 2 Activity Associated with an NQO2 Exon 3 Single-Nucleotide Polymorphism Clin. Cancer Res., March 1, 2007; 13(5): 1584 - 1590. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. Bentle, K. E. Reinicke, E. A. Bey, D. R. Spitz, and D. A. Boothman Calcium-dependent Modulation of Poly(ADP-ribose) Polymerase-1 Alters Cellular Metabolism and DNA Repair J. Biol. Chem., November 3, 2006; 281(44): 33684 - 33696. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Fraser, J. K.J. Diss, A.-M. Chioni, M. E. Mycielska, H. Pan, R. F. Yamaci, F. Pani, Z. Siwy, M. Krasowska, Z. Grzywna, et al. Voltage-Gated Sodium Channel Expression and Potentiation of Human Breast Cancer Metastasis Clin. Cancer Res., August 1, 2005; 11(15): 5381 - 5389. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. H. Ward, S. Danson, A. T. McGown, M. Ranson, N. A. Coe, G. C. Jayson, J. Cummings, R. H.J. Hargreaves, and J. Butler Preclinical Evaluation of the Pharmacodynamic Properties of 2,5-Diaziridinyl-3-Hydroxymethyl-6-Methyl-1,4-Benzoquinone Clin. Cancer Res., April 1, 2005; 11(7): 2695 - 2701. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Zhang, A. Braun, Z. Bauman, H. Olteanu, P. Madzelan, and R. Banerjee Expression Profiling of Homocysteine Junction Enzymes in the NCI60 Panel of Human Cancer Cell Lines Cancer Res., February 15, 2005; 65(4): 1554 - 1560. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. L. Dehn, S. L. Winski, and D. Ross Development of a New Isogenic Cell-Xenograft System for Evaluation of NAD(P)H:Quinone Oxidoreductase-Directed Antitumor Quinones: Evaluation of the Activity of RH1 Clin. Cancer Res., May 1, 2004; 10(9): 3147 - 3155. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Ramji, C. Lee, T. Inaba, A. V. Patterson, and D. S. Riddick Human NADPH-Cytochrome P450 Reductase Overexpression Does Not Enhance the Aerobic Cytotoxicity of Doxorubicin in Human Breast Cancer Cell Lines Cancer Res., October 15, 2003; 63(20): 6914 - 6919. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. J. Cullen, M. M. Hinkhouse, M. Grady, A. W. Gaut, J. Liu, Y. P. Zhang, C. J. Darby Weydert, F. E. Domann, and L. W. Oberley Dicumarol Inhibition of NADPH:Quinone Oxidoreductase Induces Growth Inhibition of Pancreatic Cancer via a Superoxide-mediated Mechanism Cancer Res., September 1, 2003; 63(17): 5513 - 5520. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. L. Cowen, A. V. Patterson, B. A. Telfer, R. E. Airley, S. Hobbs, R. M. Phillips, M. Jaffar, I. J. Stratford, and K. J. Williams Viral delivery of P450 reductase recapitulates the ability of constitutive overexpression of reductase enzymes to potentiate the activity of mitomycin C in human breast cancer xenografts Mol. Cancer Ther., September 1, 2003; 2(9): 901 - 909. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. L. Gustafson, D. Siegel, J. C. Rastatter, A. L. Merz, J. C. Parpal, J. K. Kepa, D. Ross, and M. E. Long Kinetics of NAD(P)H:Quinone Oxidoreductase I (NQO1) Inhibition by Mitomycin C in Vitro and in Vivo J. Pharmacol. Exp. Ther., June 1, 2003; 305(3): 1079 - 1086. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. Holtz, S. Rockwell, M. Tomasz, and A. C. Sartorelli Nuclear Overexpression of NADH:Cytochrome b5 Reductase Activity Increases the Cytotoxicity of Mitomycin C (MC) and the Total Number of MC-DNA Adducts in Chinese Hamster Ovary Cells J. Biol. Chem., February 7, 2003; 278(7): 5029 - 5034. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Kuffel, J. C. Schroeder, L. J. Pobst, S. Naylor, J. M. Reid, S. H. Kaufmann, and M. M. Ames Activation of the Antitumor Agent Aminoflavone (NSC 686288) Is Mediated by Induction of Tumor Cell Cytochrome P450 1A1/1A2 Mol. Pharmacol., July 1, 2002; 62(1): 143 - 153. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Hostein, D. Robertson, F. DiStefano, P. Workman, and P. Andrew Clarke Inhibition of Signal Transduction by the Hsp90 Inhibitor 17-Allylamino-17-demethoxygeldanamycin Results in Cytostasis and Apoptosis Cancer Res., May 1, 2001; 61(10): 4003 - 4009. [Abstract] [Full Text]
|
|
|
|
|
|
Y. Gan, Y. Mo, J. E. Kalns, J. Lu, K. Danenberg, P. Danenberg, M. G. Wientjes, and J. L-S. Au Expression of DT-Diaphorase and Cytochrome P450 Reductase Correlates with Mitomycin C Activity in Human Bladder Tumors Clin. Cancer Res., May 1, 2001; 7(5): 1313 - 1319. [Abstract] [Full Text]
|
|
|
|
 |
|
 L. J. Yu, J. Matias, D. A. Scudiero, K. M. Hite, A. Monks, E. A. Sausville, and D. J. Waxman P450 Enzyme Expression Patterns in the NCI Human Tumor Cell Line Panel Drug Metab. Dispos., March 1, 2001; 29(3): 304 - 312. [Abstract] [Full Text]
|
|
|
|
|
|
R. M. Phillips, A. M. Burger, P. M. Loadman, C. M. Jarrett, D. J. Swaine, and H.-H. Fiebig Predicting Tumor Responses to Mitomycin C on the Basis of DT-Diaphorase Activity or Drug Metabolism by Tumor Homogenates: Implications for Enzyme-directed Bioreductive Drug Development Cancer Res., November 1, 2000; 60(22): 6384 - 6390. [Abstract] [Full Text]
|
|
|
|
|
|
S. Y. Sharp, L. R. Kelland, M. R. Valenti, L. A. Brunton, S. Hobbs, and P. Workman Establishment of an Isogenic Human Colon Tumor Model for NQO1 Gene Expression: Application to Investigate the Role of DT-Diaphorase in Bioreductive Drug Activation In Vitro and In Vivo Mol. Pharmacol., November 1, 2000; 58(5): 1146 - 1155. [Abstract] [Full Text]
|
|
|
|
|
|
N. Niitsu, T. Kasukabe, A. Yokoyama, J. Okabe-Kado, Y. Yamamoto-Yamaguchi, M. Umeda, and Y. Honma Anticancer Derivative of Butyric Acid (Pivalyloxymethyl Butyrate) Specifically Potentiates the Cytotoxicity of Doxorubicin and Daunorubicin through the Suppression of Microsomal Glycosidic Activity Mol. Pharmacol., July 1, 2000; 58(1): 27 - 36. [Abstract] [Full Text]
|
|
|
|
|
|
L. M. Siemankowski, J. Morreale, B. D. Butts, and M. M. Briehl Increased Tumor Necrosis Factor-{{alpha}} Sensitivity of MCF-7 Cells Transfected with NAD(P)H:Quinone Reductase Cancer Res., July 1, 2000; 60(13): 3638 - 3644. [Abstract] [Full Text]
|
|
|
|
|
|
J. J. Pink, S. M. Planchon, C. Tagliarino, M. E. Varnes, D. Siegel, and D. A. Boothman NAD(P)H:Quinone Oxidoreductase Activity Is the Principal Determinant of beta -Lapachone Cytotoxicity J. Biol. Chem., February 25, 2000; 275(8): 5416 - 5424. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. R. Kelland, S. Y. Sharp, P. M. Rogers, T. G. Myers, and P. Workman DT-Diaphorase Expression and Tumor Cell Sensitivity to 17-Allylamino,17-demethoxygeldanamycin, an Inhibitor of Heat Shock Protein 90 J Natl Cancer Inst, November 17, 1999; 91(22): 1940 - 1949. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Alvarez, R. Robey, V. Sandor, K. Nishiyama, Y. Matsumoto, K. Paull, S. Bates, and T. Fojo Using the National Cancer Institute Anticancer Drug Screen to Assess the Effect of MRP Expression on Drug Sensitivity Profiles Mol. Pharmacol., November 1, 1998; 54(5): 802 - 814. [Abstract] [Full Text]
|
|