Combined application of comprehensive analysis for DNA modification and reporter gene mutation assay to evaluate kidneys of gpt delta rats given madder color or its constituents

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• 作者：Yuji Ishii (1)
  Shinji Takasu (1)
  Ken Kuroda (1)
  Kohei Matsushita (1)
  Aki Kijima (1)
  Takehiko Nohmi (2)
  Kumiko Ogawa (1)
  Takashi Umemura (1)
  
• 关键词：DNA adduct ; Oxidative DNA damage ; gpt delta ; In vivo mutagenicity ; Madder color
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2014
• 出版时间：April 2014
• 年：2014
• 卷：406
• 期：9-10
• 页码：2467-2475
• 全文大小：344 KB
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• 作者单位：Yuji Ishii (1)
  Shinji Takasu (1)
  Ken Kuroda (1)
  Kohei Matsushita (1)
  Aki Kijima (1)
  Takehiko Nohmi (2)
  Kumiko Ogawa (1)
  Takashi Umemura (1)
  
  1. Division of Pathology, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan
  2. Biological Safety Research Center, National Institute of Health Science, 1-178-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan
  
• ISSN：1618-2650

文摘

DNA adductome analysis using liquid chromatography–tandem mass spectrometry is a promising tool to exhaustively search DNA modifications. Given that the molecular weight of chemical-specific adducts is determined by the total molecular weights of the active form and nucleotide bases, we developed a new method of comprehensive analysis for chemical-specific DNA adducts based on the principle of adductome analysis. The actual analytical mass range was 50 mass units up or down from the average molecular weight of the four DNA bases plus the molecular weight of the expected active form of the chemical. Using lucidin-3-O-primeveroside (LuP), lucidin-modified bases formed by its active form were exhaustively searched using this new method. Various DNA adducts, including Luc-N 2-dG and Luc-N 6-dA, were identified in the kidneys of rats given LuP. Together with measurement of 8-hydroxydeoxyguanosine (8-OHdG) levels, the combined application of this new method with a reporter gene mutation assay was performed to clarify renal carcinogenesis induced by madder color (MC) that includes LuP and alizarin (Alz) as constituent agents. A DNA adductome map derived from MC-treated rats was almost identical to that of LuP-treated rats, but not Alz-treated rats. Although 8-OHdG levels were elevated in MC- and Alz-treated rats, significant increases in gpt and Spi?/sup> mutant frequencies were observed only in MC- and LuP-treated rats. In addition, the spectrum of gpt mutants in MC-treated rats showed almost the same pattern as those in LuP-treated rats. The overall data suggest that LuP may be responsible for MC-induced carcinogenicity and that the proposed methodology is appropriate for exploring and understanding mechanisms of chemical carcinogenesis. Figure DNA adductome map of kidneys from F344 gpt delta rats in the control and LuP-treated groups. The peaks detected in control and LuP-treated rats are represented as black and blue spots, respectively