Summary
Epidemiological studies suggested a protective effect of certain phenotypes of polymorphic foreign-compound-metabolizing enzymes in some types of cancer. Poor metabolizers (PM) of debrisoquine 4-hydroxylase (cytochrome P-450IID6, CYP2D6) were found to be underrepresented among patients with lung cancer. Recent advances in molecular genetic characterization of CYP2D6, glutathione S-transferase (GST) class Mu, and arylamine N-acetyltransferase enabled genotypical determination of mutant alleles in lung cancer patients. Restriction fragment length polymorphism (RFLP) with a cDNA gene probe of CYP2D6 was analyzed in 79 lung cancer patients who were phenotyped with debrisoquine. Mutant alleles were detected by allele-specific polymerase chain reaction (PCR). In the same individuals, genotype of GST class Mu was analyzed by PCR and correlated with ex vivo activity of glutathione conjugation towards trans-stilbene oxide. RFLP patterns allowed discrimination between the slow and fast genotype of N-acetyltransferase as well as the heterozygotes. Three phenotypical PMs of debrisoquine (3.8%) were confirmed by PCR and RFLP. No PM could be unambiguously recognized only by RFLP patterns. The PMs were characterized by PCR and RFLP as carriers of the 29B/29B (n=1), 29A/29B (n=1), and 29A/44 (n = 1) mutant alleles. Higher debrisoquine hydroxylase activities were found in the homozygous EMs, who possess two active genes, as compared to heterozygous EMs, who have only one active gene. The patients with phenotypically impaired GST Mu activity were confirmed as such by PCR. A complete correspondence between phenotyping of N-acetyltransferase (with caffeine) and genotyping was found. The new genetic techniques proved to be powerful tools for molecular-epidemiological studies aimed at establishing host factors of cancer susceptibility.
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Abbreviations
- AFMU:
-
5-acetylamino-6-formylamino-3methyluracil
- by:
-
base pair
- cDNA:
-
complementary DNA
- CYP2D6a:
-
cytochrome P-450IID6 (debrisoquine hydroxylase)
- CYP2D6a:
-
gene locus coding for CYP2D6
- EM:
-
extensive metabolizer
- GST:
-
glutathione S-transferase
- MR:
-
metabolic ratio
- NAT:
-
arylamine N-acetyltransferase
- NATa:
-
gene locus coding for NAT
- PCR:
-
polymerase chain reaction
- PM:
-
poor metabolizer
- RFLP:
-
restriction fragment length polymorphism
- wt:
-
wild type
- 1X:
-
1-methylxanthine
References
Ayesh R, Idle JR, Ritchie JC, Crothers MJ, Hetzel MR (1984) Metabolic oxidation phenotypes as markers for susceptibility to lung cancer. Nature 312:169–170.
Benitez J, Ladero JM, Jara C, Carrillo JA, Cobaleda J, Llerena A, Vargas E, Munoz JJ (1991) Polymorphic oxidation of debrisoquine in lung cancer patients. Eur J Cancer 27(2):161–166.
Birrer MJ, Minna JD (1988) Molecular genetics of lung cancer. Semin Oncol 15:226–235.
Bladeren PJ van (1988) Formation of toxic metabolites from drugs and other xenobiotics by glutathione conjugation. Trends Pharmacol Sci 9:295–299.
Blum M, Grant DM, McBride W, Heim M, Meyer UA (1990) Human arylamine N-acetyltransferase genes: isolation, chromosomal localization, and functional expression. DNA Cell Biol 9:193–203.
Board PG (1981) Biochemical genetics of glutathione Stransferase in man. Am J Hum Genet 33:36–43.
Brockmöller J, Gross D, Kerb R, Drakoulis N, Roots I (1992) Correlation between trans-stilbene oxide glutathione conjugation activity and the deletion mutation in the glutathione S-transferase class Mu gene detected by PCR. Biochem Pharmacol 43:647–650.
Brosen K (1990) Recent developments in hepatic drug oxidation, implications for clinical pharmacokinetics. Clin Pharmacokinet 18:220–239.
Burgess EJ, Trafford JAP (1985) Acetylator phenotype in patients with lung carcinoma — a negative report. Eur J Resp Dis 67:17–19.
Caporaso NE, Tucker MA, Hoover RN, Hayes RB, Pickle LW, Issaq HJ, Muschik GM, Green-Gallo L, Buivys D, Aisner S, Resau JH, Trump BF, Tollerud D, Weston A, Harris CC (1990) Lung cancer and the debrisoquine metabolic phenotype. J Natl Cancer Inst 82:1264–1272.
Cartwright RA, Philip PA, Rogers HJ, Glashan RW (1984) Genetically determined debrisoquine oxidation capacity in bladder cancer. Carcinogenesis 5:1191–1192.
Crespi CL, Penman BW, Gelboin HV, Gonzales FJ (1991) A tobacco smoke-derived nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, is activated by multiple human cytochrome P450s including the polymorphic human cytochrome P4502D6. Carcinogenesis 12:1197–1201.
DeJong JL, Chang CM, Whang-Peng J, Knutsen T, Tu CPD (1988) The human liver glutathione S-transferase gene superfamily: expression and chromosome mapping of an Hb subunit cDNA. Nucleic Acids Res 16:8541–8554.
Drakoulis N, Beland M, Roots I (1991) Determination of arylamine N-acetyltransferase genotype in humans. Naunyn-Schmiedeberg's Arch Pharmacol 344 (Suppl):R88.
Drakoulis N, Seeger K, Gross D, Schütt S, Vasiliades C, Brockmöller J, Roots I (1991) Determination of glutathione S-transferase μ genotype in humans by PCR-amplified gene probes. In: Rolfs A, Schumacher HC, Marx P (eds) PCR topics — usage of polymerase chain reaction in genetic and infectious diseases. Springer, Berlin Heidelberg New York, pp 94–98.
Duche JC, Joanne C, Barre J, Cremoux H de, Dalphin JC, Depierre A, Brochard P, Tillement JP, Bechtel P (1991) Lack of a relationship between the polymorphism of debrisoquine oxidation and lung cancer. Br J Clin Pharmacol 31:533–536.
Eichelbaum M, Baur MP, Dengler HJ, Osikowska-Evers BO, Tieves G, Zekorn C, Rittner C (1987) Chromosomal assignment of human cytochrome P-450 (debrisoquine/ spartein type) to chromosome 22. Cytogenet Cell Genet 46:610.
Evans WE, Relling MV (1990) Xbal 16- plus 9-kilobase DNA restriction fragments identify a mutant allele for debrisoquin hydroxylase: report of a family study. Mol Pharmacol 37:639–642.
Gibbs AR, Seal RM (1984) The histological varieties of bronchial carcinoma. In: Bates M (ed) Bronchial carcinoma: an integrated approach to diagnosis and management. Springer, Berlin Heidelberg New York, pp 129–145.
Gonzalez FJ, Skoda RC, Kimura S, Umeno M, Zanger UM, Nebert DW, Gelboin HV, Hardwick JP, Meyer UA (1988) Characterization of the common genetic defect in humans deficient in debrisoquine metabolism. Nature 331:442–446.
Gough AC, Miles JS, Spurr NK, Moss JE, Gaedigk A, Eichelbaum M, Wolf CR (1990) Identification of the primary gene defect at the cytochrome P450 CYP2D locus. Nature 347:773–775.
Grant DM, Tang BK, Kalow W (1983) Variability in caffeine metabolism. Clin Pharmacol Ther 33:591–602.
Grant DM, Blum M, Demierre A, Meyer UA (1989) Nucleotide sequence of an intronless gene for a human arylamine N-acetyltransferase related to polymorphic drug acetylation. Nucleic Acids Res 17:3978.
Guengerich FP (1988) Roles of cytochrome P-450 enzymes in chemical carcinogenesis and cancer chemotherapy. Cancer Res 48:2946–2954.
Harris CC (1989) Interindividual variation among humans in carcinogen metabolism, DNA adduct formation and DNA repair. Carcinogenesis 10:1563–1566.
Heighway J, Thatcher N, Cerny T, Hasleton PS (1986) Genetic predisposition to human lung cancer. Br J Cancer 53:453–457.
Heim M, Meyer UA (1990) Genotyping of poor metabolisers of debrisoquine by allele-specific PCR amplification. Lancet 336:529–532.
Hinson JA, Kadlubar FF (1988) Glutathione and glutathione transferases in the detoxification of drug and carcinogen metabolites. In: Sies H, Ketterer B (eds) Glutathione conjugation: mechanisms and biological significance. Academic Press, New York, pp 235–280.
Hsu IC, Metcalf RA, Sun T, Welsh JA, Wang NJ, Harris CC (1991) Mutational hotspot in the p53 gene in human hepatocellular carcinomas. Nature 350:427–428.
Hussey AJ, Hayes JD, Beckett GJ (1987) The polymorphic expression of neutral glutathione S-transferase in human mononuclear leucocytes as measured by specific radioimmunoassay. Biochem Pharmacol 36:4013–4015.
Kagimoto M, Heim M, Kagimoto K, Zeugin T, Meyer UA (1990) Multiple mutations of the human cytochrome P450IID6 gene (CYP2D6) in poor metabolizers of debrisoquine: study of the functional significance of individual mutations by expression of chimeric genes. J Biol Chem 265:17209–17214.
Kaisary A, Smith P, Jaczq E, McAllister CB, Wilkinson GR, Ray WA, Branch RA (1987) Genetic predisposition to bladder cancer, ability to hydroxylate debrisoquine and mephenytoin as risk factors. Cancer Res 47:5488–5493.
Kaplan JC, Emanuel BS (1989) Report of the committee on the genetic constitution of chromosome 22. Cytogenet Cell Genet 51:372–383.
Kimura S, Umeno M, Skoda RC, Meyer UA, Gonzalez FJ (1989) The human debrisoquine 4-hydroxylase (CYP2D) locus: sequence and identification of the polymorphic CYP2D6 gene, a related gene, and a pseudogene. Am J Hum Genet 45:889–904.
Koob M, Dekant W (1991) Bioactivation of xenobiotics by formation of toxic glutathione conjugates. Chem Biol Interactions 77:107–136.
Lai HCJ, Quin B, Grove G, Tu C-PD (1988) Gene expression of rat glutathione S-transferases. J Biol Chem 263:11389–11395.
Law MR (1990) Genetic predisposition to lung cancer. Br J Cancer 61:195–206.
Law MR, Hetzel MR, Idle JR (1989) Debrisoquine metabolism and genetic predisposition to lung cancer. Br J Cancer 59:686–687.
Lutz WK (1990) Endogenous genotoxic agents and processes as a basis of spontaneous carcinogenesis. Mutat Res 238:287–295.
Mahgoub A, Idle JR, Dring LG, Lancaster R, Smith RL (1977) Polymorphic hydroxylation of debrisoquine in man. Lancet 11:584–586.
Müller N, Brockmöller J, Roots I (1991) Extremely long plasma half-life of amitriptyline in a woman with the cytochrome P450IID6 29/29-kilobase wild-type allele - a slowly reversible interaction with fluoxetine. Ther Drug Monit 13:533–536.
Neal GE, Moss EJ, Manson MM (1988) Glutathione conjugation in oncogenesis. In: Sies H, Ketterer B (eds) Glutathione conjugation: mechanisms and biological significance. Academic Press, New York, pp 281–314.
Nebert DW (1990) Growth signal pathways. Nature 347:709–710.
Ohsako S, Deguchi T (1990) Cloning and expression of cDNAs for polymorphic and monomorphic arylamine N-acetyltransferases from human liver. J Biol Chem 265:4630–4634.
Ooi WL, Elston RC, Chen VW, Bailey-Wilson JE, Rothschild H (1986) Increased familial risk for lung cancer. J Natl Cancer Inst 76:217–222.
Preston-Martin S (1991) Evaluation of the evidence that tobacco-specific nitrosamines (TSNA) cause cancer in humans. Toxicology 21:295–298.
Price Evans DA (1989) N-Acetyltransferase. Pharmacol Ther 42:167–234.
Robertson IGC, Guthenberg C, Mannervik B, Jernström B (1986) Differences in stereoselectivity and catalytic efficiency of three human glutathione transferases in the conjugation of glutathione with 7β,8α-dihydroxy-9α,10α-oxy-7,8,9,10-tetrahydrobenzo(a)pyrene. Cancer Res 46:2220–2224.
Roots I, Drakoulis N, Brockmöller J (1992) Polymorphic enzymes and cancer risk: concepts, methodology and data review. In: Kalow W (ed) Pharmacogenetics of drug metabolism. Pergamon Press, London (International encyclopedia of pharmacology and therapeutics) (in press).
Roots I, Drakoulis N, Ploch M, Heinemeyer G, Loddenkemper R, Minks T, Nitz M, Otte F, Koch M (1988) Debrisoquine hydroxylation phenotype, acetylation phenotype, and ABO blood groups as genetic host factors of lung cancer risk. Klin Wochenschr 66 (Suppl XI): 87–97.
Roots I, Drakoulis N, Brockmöller J, Janicke I, Cuprunov M, Ritter J (1989) Hydroxylation and acetylation phenotypes as genetic risk factors in certain malignancies. In: Kato R, Estabrook RW, Cayen MN (eds) Xenobiotic metabolism and disposition. Taylor & Francis, London, pp 499–506.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Seidegard J, Pero RW (1985) The hereditary transmission of high glutathione transferase activity towards trans-stilbene oxide in human mononuclear leukocytes. Hum Genet 69:66–68.
Seidegard J, DePierre, Pero RW (1985) Hereditary interindividual differences in the glutathione transferase activity towards trans-stilbene oxide in resting human mononuclear leukocytes are due to a particular isozyme(s). Carcinogenesis 6:1211–1216.
Seidegard J, Vorachek WR, Pero RW, Pearson WR (1988) Hereditary differences in the expression of the human glutathione transferase active on trans-stilbene oxide are due to a gene deletion. Proc Natl Acad Sci USA 85:7293–7297.
Seidegard J, Pero RW, Markowitz MM, Roush G, Miller DG, Beattie EJ (1990) Isoenzyme(s) of glutathione transferase (class Mu) as a marker for the susceptibility to lung cancer: a follow up study. Carcinogenesis 11:33–36.
Sellers TA, Bailey-Wilson JE, Elston RC, Wilson AF, Elston GZ, Ooi WL, Rothschild H (1990) Evidence for mendelian inheritance in the pathogenesis of lung cancer. J Natl Cancer Inst 82:1272–1279.
Shea TC, Claflin G, Comstock KE, Sanderson BJS, Burstein NA, Keenan EJ, Mannervik B, Henner WD (1990) Glutathione transferase activity and isoenzyme composition in primary human breast cancers. Cancer Res 50:6848–6853.
Skoda RC, Gonzalez FJ, Demierre A, Meyer UA (1988) Two mutant alleles of the human cytochrome P-450db1 gene (P450C2DI) associated with genetically deficient metabolism of debrisoquine and other drugs. Proc Natl Acad Sci USA 85:5240–5243.
Speirs CJ, Murray S, Davies DS, Biola Mabadeje AF, Boohis AR (1990) Debrisoquine oxidation phenotype and susceptibility to lung cancer. Br J Clin Pharmacol 29:101–109.
Sugimura H, Caporaso NE, Shaw GL, Modali RV, Gonzalez FJ, Hoover RN, Resau JH, Trump BF, Weston A, Harris CC (1990) Human debrisoquine hydroxylase gene polymorphisms in cancer patients and controls. Carcinogenesis 11:1527–1530.
Taylor JB, Oliver J, Sherrington R, Pemble SE (1991) Structure of human glutathione S-transferase class Mu genes. Biochem J 274:587–593.
Vineis P, Simonato L (1991) Proportion of lung and bladder cancers in males resulting from occupation: a systematic approach. Arch Environ Health 46,1:6–15.
Vos RME, van Bladeren PJ (1990) Glutathione S-transferases in relation to their role in the biotransformation of xenobiotics. Chem Biol Interact 75:241–265.
Warholm M, Guthenberg C, Mannervik B, von Bahr C (1981) Purification of a new glutathione S-transferase (transferase μ) from human liver having high activity with benzo(a)pyrene-4,5-oxide. Biochem Biophys Res Commun 98:512–519.
Weston A, Willey JC, Modali R, Sugimura H, McDowell EM, Resau J, Light B, Haugen A, Mann DL, Trump BF, Harris CC (1989) Differential DNA sequence deletions from chromosomes 3, 11, 13, and 17 in squamous-cell carcinoma, large-cell carcinoma, and adenocarcinoma of the human lung. Proc Natl Acad Sci USA 86:5099–5103.
Widerstein M, Pearson WR, Engström A, Mannervik B (1991) Heterologous expression of the allelic variant Muclass glutathione transferases μ and ψ Biochem J 276:519–524.
Yue QY, Bertilsson L, Dahl-Puustinen ML, Säwe J, Sjögvist F, Johansson I, Ingelman-Sundberg M (1989) Disassociation between debrisoquine hydroxylation phenotype and genotype among Chinese. Lancet 2:870.
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Roots, I., Brockmöller, J., Drakoulis, N. et al. Mutant genes of cytochrome P-450IID6, glutathione S-transferase class Mu, and arylamine N-acetyltransferase in lung cancer patients. Clin Investig 70, 307–319 (1992). https://doi.org/10.1007/BF00184667
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DOI: https://doi.org/10.1007/BF00184667