Short-term detection of gastric genotoxicity using the DNA double-strand break marker γ-H2AX

Asako Okabe, Yuka Kiriyama, Shugo Suzuki, Kohei Sakurai, Atsushi Teramoto, Hiroyuki Kato, Aya Naiki-Ito, Sayumi Tahara, Satoru Takahashi, Makoto Kuroda, Atsushi Sugioka, Tetsuya Tsukamoto

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

DNA damage caused by Helicobacter pylori infection and chronic inflammation or exposure to genotoxic agents is considered an important risk factor of gastric carcinogenesis. In this study, we have evaluated a short-term technique to detect DNA damage response to various chemical carcinogens; it involves visualization of Ser 139-phosphorylated histone H2AX (γ-H2AX) foci by immunohistochemistry and expression analysis of other genes by quantitative RT-PCR. Six-week-old male rats were intragastrically administered N-methyl-N-nitrosourea (MNU), 3,2'-dimethyl-4-aminobiphenyl (DMAB), dimethylnitrosamine (DMN), and 1,2- dimethylhydrazine (DMH) for 5 days/week for 4 weeks, using corn oil as a vehicle. Animals were sacrificed at day 28, and their stomachs were excised. γ-H2AX foci formation, indicating DNA double-strand breaks, was observed in the proliferative zone of both fundic and pyloric glands. The number of positive cells per gland was significantly high in pyloric glands in the MNU group and in fundic glands in the MNU and DMAB groups. A significant increase in p21waf1 mRNA level was observed in the DMN group compared with the control, which was in contrast to the decreasing tendency of the h2afx mRNA level in the MNU and DMN groups. Apoptotic cells positive for γ-H2AX pan or peripheral nuclear staining were observed on the surface layer of the fundic mucosa in the MNU group. The fundic pepsinogen a5 (pga5) mRNA level showed a significant decrease, indicating gland damage. The pyloric pepsinogen c mRNA level showed no change. In conclusion, γ-H2AX in combination with other gene expression analyses could be a useful biomarker in a short-term experiment on gastric chemical genotoxicity.

Original languageEnglish
Pages (from-to)91-99
Number of pages9
JournalJournal of Toxicologic Pathology
Volume32
Issue number2
DOIs
Publication statusPublished - 01-01-2019

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Double-Stranded DNA Breaks
Dimethylnitrosamine
Stomach
Pepsinogen A
Messenger RNA
DNA
Gastric Mucosa
DNA Damage
Cells
1,2-Dimethylhydrazine
Methylnitrosourea
Corn Oil
Helicobacter Infections
Biomarkers
Gene expression
Helicobacter pylori
Carcinogens
Histones
Rats
Carcinogenesis

All Science Journal Classification (ASJC) codes

  • Pathology and Forensic Medicine
  • Toxicology

Cite this

Okabe, Asako ; Kiriyama, Yuka ; Suzuki, Shugo ; Sakurai, Kohei ; Teramoto, Atsushi ; Kato, Hiroyuki ; Naiki-Ito, Aya ; Tahara, Sayumi ; Takahashi, Satoru ; Kuroda, Makoto ; Sugioka, Atsushi ; Tsukamoto, Tetsuya. / Short-term detection of gastric genotoxicity using the DNA double-strand break marker γ-H2AX. In: Journal of Toxicologic Pathology. 2019 ; Vol. 32, No. 2. pp. 91-99.
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abstract = "DNA damage caused by Helicobacter pylori infection and chronic inflammation or exposure to genotoxic agents is considered an important risk factor of gastric carcinogenesis. In this study, we have evaluated a short-term technique to detect DNA damage response to various chemical carcinogens; it involves visualization of Ser 139-phosphorylated histone H2AX (γ-H2AX) foci by immunohistochemistry and expression analysis of other genes by quantitative RT-PCR. Six-week-old male rats were intragastrically administered N-methyl-N-nitrosourea (MNU), 3,2'-dimethyl-4-aminobiphenyl (DMAB), dimethylnitrosamine (DMN), and 1,2- dimethylhydrazine (DMH) for 5 days/week for 4 weeks, using corn oil as a vehicle. Animals were sacrificed at day 28, and their stomachs were excised. γ-H2AX foci formation, indicating DNA double-strand breaks, was observed in the proliferative zone of both fundic and pyloric glands. The number of positive cells per gland was significantly high in pyloric glands in the MNU group and in fundic glands in the MNU and DMAB groups. A significant increase in p21waf1 mRNA level was observed in the DMN group compared with the control, which was in contrast to the decreasing tendency of the h2afx mRNA level in the MNU and DMN groups. Apoptotic cells positive for γ-H2AX pan or peripheral nuclear staining were observed on the surface layer of the fundic mucosa in the MNU group. The fundic pepsinogen a5 (pga5) mRNA level showed a significant decrease, indicating gland damage. The pyloric pepsinogen c mRNA level showed no change. In conclusion, γ-H2AX in combination with other gene expression analyses could be a useful biomarker in a short-term experiment on gastric chemical genotoxicity.",
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Okabe, A, Kiriyama, Y, Suzuki, S, Sakurai, K, Teramoto, A, Kato, H, Naiki-Ito, A, Tahara, S, Takahashi, S, Kuroda, M, Sugioka, A & Tsukamoto, T 2019, 'Short-term detection of gastric genotoxicity using the DNA double-strand break marker γ-H2AX', Journal of Toxicologic Pathology, vol. 32, no. 2, pp. 91-99. https://doi.org/10.1293/tox.2019-0007

Short-term detection of gastric genotoxicity using the DNA double-strand break marker γ-H2AX. / Okabe, Asako; Kiriyama, Yuka; Suzuki, Shugo; Sakurai, Kohei; Teramoto, Atsushi; Kato, Hiroyuki; Naiki-Ito, Aya; Tahara, Sayumi; Takahashi, Satoru; Kuroda, Makoto; Sugioka, Atsushi; Tsukamoto, Tetsuya.

In: Journal of Toxicologic Pathology, Vol. 32, No. 2, 01.01.2019, p. 91-99.

Research output: Contribution to journalArticle

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T1 - Short-term detection of gastric genotoxicity using the DNA double-strand break marker γ-H2AX

AU - Okabe, Asako

AU - Kiriyama, Yuka

AU - Suzuki, Shugo

AU - Sakurai, Kohei

AU - Teramoto, Atsushi

AU - Kato, Hiroyuki

AU - Naiki-Ito, Aya

AU - Tahara, Sayumi

AU - Takahashi, Satoru

AU - Kuroda, Makoto

AU - Sugioka, Atsushi

AU - Tsukamoto, Tetsuya

PY - 2019/1/1

Y1 - 2019/1/1

N2 - DNA damage caused by Helicobacter pylori infection and chronic inflammation or exposure to genotoxic agents is considered an important risk factor of gastric carcinogenesis. In this study, we have evaluated a short-term technique to detect DNA damage response to various chemical carcinogens; it involves visualization of Ser 139-phosphorylated histone H2AX (γ-H2AX) foci by immunohistochemistry and expression analysis of other genes by quantitative RT-PCR. Six-week-old male rats were intragastrically administered N-methyl-N-nitrosourea (MNU), 3,2'-dimethyl-4-aminobiphenyl (DMAB), dimethylnitrosamine (DMN), and 1,2- dimethylhydrazine (DMH) for 5 days/week for 4 weeks, using corn oil as a vehicle. Animals were sacrificed at day 28, and their stomachs were excised. γ-H2AX foci formation, indicating DNA double-strand breaks, was observed in the proliferative zone of both fundic and pyloric glands. The number of positive cells per gland was significantly high in pyloric glands in the MNU group and in fundic glands in the MNU and DMAB groups. A significant increase in p21waf1 mRNA level was observed in the DMN group compared with the control, which was in contrast to the decreasing tendency of the h2afx mRNA level in the MNU and DMN groups. Apoptotic cells positive for γ-H2AX pan or peripheral nuclear staining were observed on the surface layer of the fundic mucosa in the MNU group. The fundic pepsinogen a5 (pga5) mRNA level showed a significant decrease, indicating gland damage. The pyloric pepsinogen c mRNA level showed no change. In conclusion, γ-H2AX in combination with other gene expression analyses could be a useful biomarker in a short-term experiment on gastric chemical genotoxicity.

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