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The application of hepatic P450 reductase null gpt delta mice in studying the role of hepatic P450 in genotoxic carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced mutagenesis

  • Genotoxicity and Carcinogenicity
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Abstract

The cytochrome P450 (P450 or CYP) is involved in both detoxification and metabolic activation of many carcinogens. In order to identify the role of hepatic P450 in the mutagenesis of genotoxic carcinogens, we generated a novel hepatic P450 reductase null (HRN) gpt delta mouse model, which lacks functional hepatic P450 on a gpt delta mouse background. In this study, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) was used to treat HRN gpt delta mice and control littermates. Gene mutations in the liver and lungs were detected, and mutation spectra were analyzed. Pharmacokinetic analyses were performed, and tissue levels of NNK and metabolite were determined. NNK-induced mutant frequencies (MFs) were equivalent to spontaneous MFs in the liver, but increased more than 3 times in the lungs of HRN gpt delta mice compared to control mice. NNK-induced mutation spectra showed no difference between HRN gpt delta mice and control littermates. Toxicokinetic studies revealed reduced clearance of NNK with elevated tissue concentrations in HRN gpt delta mice. To our knowledge, these are the first data demonstrating that NNK cannot induce mutagenesis in the liver without P450 metabolic activation, but can induce mutagenesis in lungs by a hepatic P450-independent mechanism. Moreover, our data show that hepatic P450 plays a major role in the systemic clearance of NNK, thereby protecting the lungs against NNK-induced mutagenesis. Our model will be useful in establishing the role of hepatic versus extrahepatic P450-mediated mutagenesis, and the relative contributions of P450 compared to other biotransformation enzymes in the genotoxic carcinogens’ activation.

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Acknowledgments

This work was supported by the National Natural Scientific Foundation of China [21077112] and Key Projects of National Science and the Technology Pillar Program [2008ZX09305-007, 2009ZX09501-033]. We thank Dr. Tao Chen (NCTR, USA) for helpful discussions, comments, and criticisms.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Center for Drug Safety and Evaluation Research, State Key Laboratory of New Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Haike road 501, Shanghai, 201203, China

    Yang Luan, Guozhen Xing, Xinming Qi, Mengjun Wu, Chenggang Li, Jun Yao, Likun Gong & Jin Ren

  2. Graduate School of the Chinese Academy of Sciences, Shanghai, 201203, China

    Mengjun Wu, Chenggang Li & Jun Yao

  3. Division of Genetics and Mutagenesis, National Institute of Health Sciences, Tokyo, 158-8501, Japan

    Takehiko Nohmi

  4. New York State Department of Health, Wadsworth Center, Albany, NY, 12201-0509, USA

    Jun Gu

  5. School of Public Health, State University of New York at Albany, Albany, NY, 12201-0509, USA

    Jun Gu

  6. Shanghai Tobacco Group Co., Ltd., Shanghai, 200082, China

    Wanhong Zhou & Saijing Zheng

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  1. Yang Luan
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  2. Guozhen Xing
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Correspondence to Yang Luan or Jin Ren.

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Yang Luan and Guozhen Xing contributed equally to this work.

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204_2012_891_MOESM1_ESM.tif

Supplementary Fig. 1 Histological examination of livers from 2-month-old female HRN gpt delta transgenic mice and control littermates. A. Liver of a control littermates; centrilobular hepatocytes have fragmented, glycogen-rich cytoplasm. B. Liver of HRN gpt delta transgenic mice; midzonal hepatocytes have large and small clear vacuoles typical of micro- and macrovesicular hepatic lipidosis. (TIFF 6330 kb)

Supplementary Table 1 Mutant Frequency of the gpt gene in the lungs of mice treated with NNK. (DOC 49 kb)

Supplementary Table 2 Mutant Frequency of the gpt gene in the liver of mice treated with NNK. (DOC 51 kb)

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Luan, Y., Xing, G., Qi, X. et al. The application of hepatic P450 reductase null gpt delta mice in studying the role of hepatic P450 in genotoxic carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced mutagenesis. Arch Toxicol 86, 1753–1761 (2012). https://doi.org/10.1007/s00204-012-0891-6

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