This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Belinsky, S. A. Articles by Maronpot, R. R. Search for Related Content PubMed Articles by Belinsky, S. A. Articles by Maronpot, R. R.

Dose-Response Relationship between O⁶-Methylguanine Formation in Clara Cells and Induction of Pulmonary Neoplasia in the Rat by 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone

Laboratory of Molecular Toxicology [S. A. B., C. M. W., M. W. A.] and Chemical Pathology Branch [J. F. F., R. R. M.], National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709

The relationship between the formation of O⁶-methylguanine (O⁶MG) and the induction of lung, liver, and nasal tumors in the Fischer 344 rat by the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) was examined in a dose-response study. Animals were treated for 20 wk (3 times/wk) with concentrations of NNK ranging from 0.03 to 50 mg/kg to induce tumors. Steady-state concentrations of O⁶MG were quantitated, and cytotoxicity was assessed in target cells and tissues after 4 wk of treatment with NNK. No cytotoxicity was detected in the lung during treatment with NNK. The formation of O⁶MG was greatest in Clara cells compared with macrophages, type II cells, small cells, and whole lung at all doses examined. The difference in adduct concentration between the Clara cell and other pulmonary cell types was most pronounced with low doses of carcinogen. The O⁶MG:dose ratio, an index of alkylation efficiency, increased 29-fold as the dose of NNK was decreased from 50 to 1 mg/kg of carcinogen. In contrast, only a small increase in alkylation efficiency was observed in type II cells and whole lung. A significant number of tumors were induced in the lung at doses of 0.1 to 50 mg/kg with incidences ranging from 10% at the lowest dose up to 87% in the group of animals which received 50 mg/kg of NNK. A linear relationship was observed when the concentration of O⁶MG in Clara cells as a function of dose was plotted against the corresponding tumor incidence. This relationship was not observed using DNA adduct concentrations in type II cells or whole lung. The development of pulmonary tumors appeared to involve the formation of alveolar hyperplasias which progressed to adenomas and finally to carcinomas. The majority of adenomas were solid, whereas carcinomas were mainly papillary. Examination of the ultrastructure of the hyperplasias, adenomas, and carcinomas revealed morphological structures (e.g., lamellar bodies, tubular myelin) which are associated with type II cells. Thus, these data suggest that the majority of neoplasms in the lung begin as type II cell proliferations with progression to adenomas and carcinomas within the areas of hyperplasia. The lack of agreement between biochemical and morphological findings makes it difficult to hypothesize a cell of origin for the pulmonary neoplasms. In contrast to the lung, tumors were induced in the liver and nasal passages only after exposure to high doses of NNK. Moreover, both the formation of DNA adducts and cytotoxicity appear obligatory for the generation of tumors in these tissues. These studies indicate that the concentration of O⁶MG in the Clara cell is an excellent indicator of the carcinogenic potency of NNK in the lung and, based on the biochemical data, also support the involvement of the Clara cell in the development of pulmonary neoplasms induced by NNK in the rat. In addition, these studies support the premise that the quantitation of DNA damage in target tissues at doses below and inclusive of standard carcinogenicity studies should aid in the estimation of risk from exposure to environmental carcinogens.

¹ To whom requests for reprints should be addressed.

Received 11/27/89. Revised 2/26/90.

This article has been cited by other articles:

				P. Upadhyaya, B. R. Lindgren, and S. S. Hecht Comparative Levels of O6-Methylguanine, Pyridyloxobutyl-, and Pyridylhydroxybutyl-DNA Adducts in Lung and Liver of Rats Treated Chronically with the Tobacco-Specific Carcinogen 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone Drug Metab. Dispos., June 1, 2009; 37(6): 1147 - 1151. [Abstract] [Full Text] [PDF]

				T. R. Church, K. E. Anderson, N. E. Caporaso, M. S. Geisser, C. T. Le, Y. Zhang, A. R. Benoit, S. G. Carmella, and S. S. Hecht A Prospectively Measured Serum Biomarker for a Tobacco-Specific Carcinogen and Lung Cancer in Smokers Cancer Epidemiol. Biomarkers Prev., January 1, 2009; 18(1): 260 - 266. [Abstract] [Full Text] [PDF]

				C. J. Gallagher, J. E. Muscat, A. N. Hicks, Y. Zheng, A.-M. Dyer, G. A. Chase, J. Richie, and P. Lazarus The UDP-Glucuronosyltransferase 2B17 Gene Deletion Polymorphism: Sex-Specific Association with Urinary 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanol Glucuronidation Phenotype and Risk for Lung Cancer Cancer Epidemiol. Biomarkers Prev., April 1, 2007; 16(4): 823 - 828. [Abstract] [Full Text] [PDF]

				A. M. Jeffrey, M. J. Iatropoulos, and G. M. Williams Nasal Cytotoxic and Carcinogenic Activities of Systemically Distributed Organic Chemicals Toxicol Pathol, December 1, 2006; 34(7): 827 - 852. [Abstract] [Full Text] [PDF]

				L. C. Pulling, B. R. Vuillemenot, J. A. Hutt, T. R. Devereux, and S. A. Belinsky Aberrant Promoter Hypermethylation of the Death-Associated Protein Kinase Gene Is Early and Frequent in Murine Lung Tumors Induced by Cigarette Smoke and Tobacco Carcinogens Cancer Res., June 1, 2004; 64(11): 3844 - 3848. [Abstract] [Full Text] [PDF]

				D. Wiener, J.-L. Fang, N. Dossett, and P. Lazarus Correlation between UDP-Glucuronosyltransferase Genotypes and 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanone Glucuronidation Phenotype in Human Liver Microsomes Cancer Res., February 1, 2004; 64(3): 1190 - 1196. [Abstract] [Full Text] [PDF]

				S. S. Hecht, S. E. Murphy, S. G. Carmella, C. L. Zimmerman, L. Losey, I. Kramarczuk, M. R. Roe, S. S. Puumala, Y. S. Li, C. Le, et al. Effects of Reduced Cigarette Smoking on the Uptake of a Tobacco-Specific Lung Carcinogen J Natl Cancer Inst, January 21, 2004; 96(2): 107 - 115. [Abstract] [Full Text] [PDF]

				D. Wiener, D. R. Doerge, J.-L. Fang, P. Upadhyaya, and P. Lazarus CHARACTERIZATION OF N-GLUCURONIDATION OF THE LUNG CARCINOGEN 4-(METHYLNITROSAMINO)-1-(3-PYRIDYL)-1-BUTANOL (NNAL) IN HUMAN LIVER: IMPORTANCE OF UDP-GLUCURONOSYLTRANSFERASE 1A4 Drug Metab. Dispos., January 1, 2004; 32(1): 72 - 79. [Abstract] [Full Text] [PDF]

				E. Piras, A. Franzen, E. L. Fernandez, U. Bergstrom, F. Raffalli-Mathieu, M. Lang, and E. B. Brittebo Cell-specific Expression of CYP2A5 in the Mouse Respiratory Tract: Effects of Olfactory Toxicants J. Histochem. Cytochem., November 1, 2003; 51(11): 1545 - 1555. [Abstract] [Full Text] [PDF]

				Y. Tesfaigzi, P. S. Wright, and S. A. Belinsky SPRR1B overexpression enhances entry of cells into the G0 phase of the cell cycle Am J Physiol Lung Cell Mol Physiol, October 1, 2003; 285(4): L889 - L898. [Abstract] [Full Text] [PDF]

				G. Boysen, P. M.J. Kenney, P. Upadhyaya, M. Wang, and S. S. Hecht Effects of benzyl isothiocyanate and 2-phenethyl isothiocyanate on benzo[a]pyrene and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone metabolism in F-344 rats Carcinogenesis, March 1, 2003; 24(3): 517 - 525. [Abstract] [Full Text] [PDF]

				P. E. Jackson, P. J. O'Connor, D. P. Cooper, G. P. Margison, and A. C. Povey Associations between tissue-specific DNA alkylation, DNA repair and cell proliferation in the colon and colon tumour yield in mice treated with 1,2-dimethylhydrazine Carcinogenesis, March 1, 2003; 24(3): 527 - 533. [Abstract] [Full Text] [PDF]

				S. S. Hecht Human urinary carcinogen metabolites: biomarkers for investigating tobacco and cancer Carcinogenesis, June 1, 2002; 23(6): 907 - 922. [Abstract] [Full Text] [PDF]

				S. S. Hecht, S. G. Carmella, M. Ye, K.-a. Le, J. A. Jensen, C. L. Zimmerman, and D. K. Hatsukami Quantitation of Metabolites of 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone after Cessation of Smokeless Tobacco Use Cancer Res., January 1, 2002; 62(1): 129 - 134. [Abstract] [Full Text] [PDF]

				Z. Wu, P. Upadhyaya, S. G. Carmella, S. S. Hecht, and C. L. Zimmerman Disposition of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in bile duct-cannulated rats: Stereoselective metabolism and tissue distribution Carcinogenesis, January 1, 2002; 23(1): 171 - 179. [Abstract] [Full Text] [PDF]

				L. A. Peterson, N. M. Thomson, D. L. Crankshaw, E. E. Donaldson, and P. J. Kenney Interactions between Methylating and Pyridyloxobutylating Agents in A/J Mouse Lungs: Implications for 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced Lung Tumorigenesis Cancer Res., August 1, 2001; 61(15): 5757 - 5763. [Abstract] [Full Text] [PDF]

				L. C. Pulling, D. M. Klinge, and S. A. Belinsky p16INK4a and {beta}-catenin alterations in rat liver tumors induced by NNK Carcinogenesis, March 1, 2001; 22(3): 461 - 466. [Abstract] [Full Text] [PDF]

				Q. Ren, S. E. Murphy, Z. Zheng, and P. Lazarus O-Glucuronidation of the Lung Carcinogen 4-(methylnitrosamino)-1- (3-Pyridyl)-1-Butanol (nnal) by Human Udp-Glucuronosyltransferases 2b7 and 1a9 Drug Metab. Dispos., November 1, 2000; 28(11): 1352 - 1360. [Abstract] [Full Text]

				A. C. Povey DNA Adducts: Endogenous and Induced Toxicol Pathol, May 1, 2000; 28(3): 405 - 414. [Abstract] [PDF]

				J. Mimoto, K. Kiura, K. Matsuo, T. Yoshino, I. Takata, H. Ueoka, M. Kataoka, and M. Harada (-)-Epigallocatechin gallate can prevent cisplatin-induced lung tumorigenesis in A/J mice Carcinogenesis, May 1, 2000; 21(5): 915 - 919. [Abstract] [Full Text] [PDF]

				E. Schrader, K. I. Hirsch-Ernst, E. Scholz, G. F. Kahl, and H. Foth Metabolism of 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in Primary Cultures of Rat Alveolar Type II Cells Drug Metab. Dispos., February 1, 2000; 28(2): 180 - 185. [Abstract] [Full Text]

				B. D. Reh, D.G. DeBord, M. A. Butler, T. M. Reid, C. Mueller, and J. M. Fajen O6-methylguanine DNA adducts associated with occupational nitrosamine exposure Carcinogenesis, January 1, 2000; 21(1): 29 - 33. [Abstract] [Full Text] [PDF]

				Q. Ren, S. E. Murphy, A. J. Dannenberg, J. Y. Park, T. R. Tephly, and P. Lazarus Glucuronidation of the Lung Carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) by Rat UDP-Glucuronosyltransferase 2B1 Drug Metab. Dispos., September 1, 1999; 27(9): 1010 - 1016. [Abstract] [Full Text]

				X. Qin, H. Zhou, L. Liu, and S. L. Gerson Transgenic expression of human MGMT blocks the hypersensitivity of PMS2-deficient mice to low dose MNU thymic lymphomagenesis Carcinogenesis, September 1, 1999; 20(9): 1667 - 1673. [Abstract] [Full Text] [PDF]

				S. A. Belinsky, K. J. Nikula, W. A. Palmisano, R. Michels, G. Saccomanno, E. Gabrielson, S. B. Baylin, and J. G. Herman Aberrant methylation of p16INK4a is an early event in lung cancer and a potential biomarker for early diagnosis PNAS, September 29, 1998; 95(20): 11891 - 11896. [Abstract] [Full Text] [PDF]

				T. E. Gram Chemically Reactive Intermediates and Pulmonary Xenobiotic Toxicity Pharmacol. Rev., December 1, 1997; 49(4): 297 - 342. [Abstract] [Full Text] [PDF]

				M. E. Staretz, S. E. Murphy, C. J. Patten, M. G. Nunes, W. Koehl, S. Amin, L. A. Koenig, F. P. Guengerich, and S. S. Hecht Comparative Metabolism of the Tobacco-Related Carcinogens Benzo[a]pyrene, 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone, 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanol, and N'-Nitrosonornicotine in Human Hepatic Microsomes Drug Metab. Dispos., February 1, 1997; 25(2): 154 - 162. [Abstract] [Full Text]

				G. A. Boorman, R. Hmley, S. Grumbein, B. J. Chou, R. A. Herbert, T. Goehl, P. W. Mellick, J. H. Roycroft, J. K. Haseman, and R. Sills Toxicology and Carcinogenesis Studies of Ozone and Ozone 4-(N-Nitrosomethylamino)-1-(3-Pyridyl)-1-Butanone in Fischer-344/N Rats Toxicol Pathol, September 1, 1994; 22(5): 545 - 554. [Abstract] [PDF]