• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of brjcancerBJC HomepageBJC Advance online publicationBJC Current IssueSubmitting an article to BJCWeb feeds
Br J Cancer. Sep 1995; 72(3): 555–561.
PMCID: PMC2033894

Presence of a heterozygous substitution and its relationship to DT-diaphorase activity.

Abstract

A point mutation in the mRNA of NADP(H): quinone oxidoreductase 1 (NQO1, DT-diaphorase) is believed to be responsible for reduced enzyme activity in the adenocarcinoma BE cell line. The present study examined nine cultured human non-cancerous fibroblast cell strains, five of which were from members of a single cancer-prone family, which demonstrated widely varying activity levels of DT-diaphorase (41 - 3462 nmol min-1 mg-1 protein), to determine if genetic alteration of the NQO1 or NOQ2 gene was involved in determining enzyme activity. All cell strains expressed NQO1 and NQO2 mRNA as measured by a quantitative polymerase chain reaction amplification technique. No relationship was found between the level of mRNA expressed and the enzyme activity in the cells. Sequencing of the entire complementary DNA from the cell strains revealed only a single base substitution at nucleotide 609 in one allele encoding NQO1 in every cell strain from members of the cancer-prone family, except for one cell strain which expressed only the T at nucleotide 609 in both alleles. Subsequent examination of genomic DNA from 44 individuals revealed that this base substitution is present in approximately 50% of the population. The presence of the T at nucleotide 609 in the NQO1 locus does not appear to be directly causal for altered DT-diaphorase activity.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.5M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Belinsky M, Jaiswal AK. NAD(P)H:quinone oxidoreductase1 (DT-diaphorase) expression in normal and tumor tissues. Cancer Metastasis Rev. 1993 Jun;12(2):103–117. [PubMed]
  • Benson AM, Hunkeler MJ, Talalay P. Increase of NAD(P)H:quinone reductase by dietary antioxidants: possible role in protection against carcinogenesis and toxicity. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5216–5220. [PMC free article] [PubMed]
  • Cresteil T, Jaiswal AK. High levels of expression of the NAD(P)H:quinone oxidoreductase (NQO1) gene in tumor cells compared to normal cells of the same origin. Biochem Pharmacol. 1991 Aug 8;42(5):1021–1027. [PubMed]
  • Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS. 'Touchdown' PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res. 1991 Jul 25;19(14):4008–4008. [PMC free article] [PubMed]
  • Edwards YH, Potter J, Hopkinson DA. Human FAD-dependent NAD(P)H diaphorase. Biochem J. 1980 May 1;187(2):429–436. [PMC free article] [PubMed]
  • Eickelmann P, Schulz WA, Rohde D, Schmitz-Dräger B, Sies H. Loss of heterozygosity at the NAD(P)H: quinone oxidoreductase locus associated with increased resistance against mitomycin C in a human bladder carcinoma cell line. Biol Chem Hoppe Seyler. 1994 Jul;375(7):439–445. [PubMed]
  • Eickelmann P, Ebert T, Warskulat U, Schulz WA, Sies H. Expression of NAD(P)H:quinone oxidoreductase and glutathione S-transferases alpha and pi in human renal cell carcinoma and in kidney cancer-derived cell lines. Carcinogenesis. 1994 Feb;15(2):219–225. [PubMed]
  • Fisher GR, Patterson LH, Gutierrez PL. A comparison of free radical formation by quinone antitumour agents in MCF-7 cells and the role of NAD(P)H (quinone-acceptor) oxidoreductase (DT-diaphorase). Chem Biol Interact. 1993 Sep;88(2-3):137–153. [PubMed]
  • Fisher GR, Gutierrez PL, Oldcorne MA, Patterson LH. NAD(P)H (quinone acceptor) oxidoreductase (DT-diaphorase)-mediated two-electron reduction of anthraquinone-based antitumour agents and generation of hydroxyl radicals. Biochem Pharmacol. 1992 Feb 4;43(3):575–585. [PubMed]
  • Fisher RA, Edwards YH, Putt W, Potter J, Hopkinson DA. An interpretation of human diaphorase isozymes in terms of three gene loci DIA1, DIA2 and DIA3. Ann Hum Genet. 1977 Oct;41(2):139–149. [PubMed]
  • Fraumeni JF, Jr, Vogel CL, Easton JM. Sarcomas and multiple polyposis in a kindred. A genetic variety of hereditary polyposis? Arch Intern Med. 1968 Jan;121(1):57–61. [PubMed]
  • Gibson NW, Hartley JA, Butler J, Siegel D, Ross D. Relationship between DT-diaphorase-mediated metabolism of a series of aziridinylbenzoquinones and DNA damage and cytotoxicity. Mol Pharmacol. 1992 Sep;42(3):531–536. [PubMed]
  • Halliwell B, Gutteridge JM. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984 Apr 1;219(1):1–14. [PMC free article] [PubMed]
  • Jaiswal AK. Human NAD(P)H:quinone oxidoreductase2. Gene structure, activity, and tissue-specific expression. J Biol Chem. 1994 May 20;269(20):14502–14508. [PubMed]
  • Jaiswal AK, McBride OW, Adesnik M, Nebert DW. Human dioxin-inducible cytosolic NAD(P)H:menadione oxidoreductase. cDNA sequence and localization of gene to chromosome 16. J Biol Chem. 1988 Sep 25;263(27):13572–13578. [PubMed]
  • Jaiswal AK, Burnett P, Adesnik M, McBride OW. 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. 1990 Feb 20;29(7):1899–1906. [PubMed]
  • Kuehl BL, Buchwald M, Rauth AM. Characterization of a set of Chinese hamster ovary variant cell lines demonstrating differing sensitivity to mitomycin C. Oncol Res. 1993;5(6-7):213–221. [PubMed]
  • Lind C, Hochstein P, Ernster L. DT-diaphorase as a quinone reductase: a cellular control device against semiquinone and superoxide radical formation. Arch Biochem Biophys. 1982 Jun;216(1):178–185. [PubMed]
  • Marshall RS, Paterson MC, Rauth AM. Deficient activation by a human cell strain leads to mitomycin resistance under aerobic but not hypoxic conditions. Br J Cancer. 1989 Mar;59(3):341–346. [PMC free article] [PubMed]
  • Marshall RS, Paterson MC, Rauth AM. DT-diaphorase activity and mitomycin C sensitivity in non-transformed cell strains derived from members of a cancer-prone family. Carcinogenesis. 1991 Jul;12(7):1175–1180. [PubMed]
  • Mirzayans R, Aubin RA, Bosnich W, Blattner WA, Paterson MC. Abnormal pattern of post-gamma-ray DNA replication in radioresistant fibroblast strains from affected members of a cancer-prone family with Li-Fraumeni syndrome. Br J Cancer. 1995 Jun;71(6):1221–1230. [PMC free article] [PubMed]
  • Nebert DW. Drug-metabolizing enzymes in ligand-modulated transcription. Biochem Pharmacol. 1994 Jan 13;47(1):25–37. [PubMed]
  • Noonan KE, Beck C, Holzmayer TA, Chin JE, Wunder JS, Andrulis IL, Gazdar AF, Willman CL, Griffith B, Von Hoff DD, et al. Quantitative analysis of MDR1 (multidrug resistance) gene expression in human tumors by polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7160–7164. [PMC free article] [PubMed]
  • Riley RJ, Workman P. DT-diaphorase and cancer chemotherapy. Biochem Pharmacol. 1992 Apr 15;43(8):1657–1669. [PubMed]
  • Ross D, Siegel D, Beall H, Prakash AS, Mulcahy RT, Gibson NW. DT-diaphorase in activation and detoxification of quinones. Bioreductive activation of mitomycin C. Cancer Metastasis Rev. 1993 Jun;12(2):83–101. [PubMed]
  • Sartorelli AC. Therapeutic attack of hypoxic cells of solid tumors: presidential address. Cancer Res. 1988 Feb 15;48(4):775–778. [PubMed]
  • Siegel D, Beall H, Senekowitsch C, Kasai M, Arai H, Gibson NW, Ross D. Bioreductive activation of mitomycin C by DT-diaphorase. Biochemistry. 1992 Sep 1;31(34):7879–7885. [PubMed]
  • Talalay P, Benson AM. Elevation of quinone reductase activity by anticarcinogenic antioxidants. Adv Enzyme Regul. 1982;20:287–300. [PubMed]
  • Thor H, Smith MT, Hartzell P, Bellomo G, Jewell SA, Orrenius S. The metabolism of menadione (2-methyl-1,4-naphthoquinone) by isolated hepatocytes. A study of the implications of oxidative stress in intact cells. J Biol Chem. 1982 Oct 25;257(20):12419–12425. [PubMed]
  • Tomasz M, Chawla AK, Lipman R. Mechanism of monofunctional and bifunctional alkylation of DNA by mitomycin C. Biochemistry. 1988 May 3;27(9):3182–3187. [PubMed]
  • Traver RD, Horikoshi T, Danenberg KD, Stadlbauer TH, Danenberg PV, Ross D, Gibson NW. NAD(P)H:quinone oxidoreductase gene expression in human colon carcinoma cells: characterization of a mutation which modulates DT-diaphorase activity and mitomycin sensitivity. Cancer Res. 1992 Feb 15;52(4):797–802. [PubMed]
  • Winship PR. An improved method for directly sequencing PCR amplified material using dimethyl sulphoxide. Nucleic Acids Res. 1989 Feb 11;17(3):1266–1266. [PMC free article] [PubMed]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...