Extensive DNA damage induced by monochloramine in gastric cells

Hidekazu Suzuki, Mikiji Mori, Masayuki Suzuki, Kazushi Sakurai, Soichiro Miura, Hiromasa Ishii

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Colonization of Helicobacter pylori (Hp) to gastric mucosa plays an important role for the pathogenesis of gastric mucosal lesions. We previously reported the importance of monochloramine (NH2Cl), which was derived from the interaction between Hp-urease and infiltrated leukocytes, in the course of Hp-associated gastric mucosal injury. While the long-term infection of Hp in the gastric mucosa is known to be one of the virulent factors which closely link to the gastric carcinogenesis, the details of its pathogenetic mechanisms remain speculative. The present study is designed to examine whether a NH2Cl could damage the DNA of gastric cells. Rabbit gastric mucosal cells (RGMC) or KATO III cells were cultured and suspended. Cell suspensions were exposed to HOCl, NH3 or NH2Cl for 15 min to give a final concentration of 0.1 mM. The magnitude of a double strand break of DNA was quantified by measuring the remnant double strand stained by ethidium bromide (EB), and the fluorescence intensity of EB was analyzed by spectrophotometer. Separately, cell nuclei were stained by fluorescent dye (Hoechst No. 33258) in order to evaluate the levels of chromatin condensation evoked by DNA fragmentation. The number of cells with chromatin condensation was counted. During the entire experimental period, more than 85% of cells were persistently viable in all groups. NH2Cl significantly induced the DNA double strand break as well as chromatin condensation in RGMC and KATO III cells (P < 0.05). However, NH3 or HOCl did not induce the DNA double strand break as well as chromatin condensation in both cells. NH2Cl, but not its precursors (NH3 or HOCl), enhanced the levels of DNA injury, suggesting the possible involvement in the carcinogenesis of Hp-associated gastric mucosa.

Original languageEnglish
Pages (from-to)243-248
Number of pages6
JournalCancer Letters
Volume115
Issue number2
DOIs
Publication statusPublished - 1997 May 19

Fingerprint

DNA Damage
Stomach
Helicobacter pylori
Chromatin
Double-Stranded DNA Breaks
Gastric Mucosa
Ethidium
Carcinogenesis
Rabbits
Bisbenzimidazole
Urease
DNA Fragmentation
chloramine
Cell Nucleus
Fluorescent Dyes
Cultured Cells
Suspensions
Leukocytes
Cell Count
Fluorescence

Keywords

  • Apoptosis
  • Chromatin condensation
  • Helicobacter pylori
  • Leukocyte
  • Urease

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Oncology

Cite this

Suzuki, H., Mori, M., Suzuki, M., Sakurai, K., Miura, S., & Ishii, H. (1997). Extensive DNA damage induced by monochloramine in gastric cells. Cancer Letters, 115(2), 243-248. https://doi.org/10.1016/S0304-3835(97)04745-9

Extensive DNA damage induced by monochloramine in gastric cells. / Suzuki, Hidekazu; Mori, Mikiji; Suzuki, Masayuki; Sakurai, Kazushi; Miura, Soichiro; Ishii, Hiromasa.

In: Cancer Letters, Vol. 115, No. 2, 19.05.1997, p. 243-248.

Research output: Contribution to journalArticle

Suzuki, H, Mori, M, Suzuki, M, Sakurai, K, Miura, S & Ishii, H 1997, 'Extensive DNA damage induced by monochloramine in gastric cells', Cancer Letters, vol. 115, no. 2, pp. 243-248. https://doi.org/10.1016/S0304-3835(97)04745-9
Suzuki H, Mori M, Suzuki M, Sakurai K, Miura S, Ishii H. Extensive DNA damage induced by monochloramine in gastric cells. Cancer Letters. 1997 May 19;115(2):243-248. https://doi.org/10.1016/S0304-3835(97)04745-9
Suzuki, Hidekazu ; Mori, Mikiji ; Suzuki, Masayuki ; Sakurai, Kazushi ; Miura, Soichiro ; Ishii, Hiromasa. / Extensive DNA damage induced by monochloramine in gastric cells. In: Cancer Letters. 1997 ; Vol. 115, No. 2. pp. 243-248.
@article{86a6c2454ccd4cb48b77ed7af3318888,
title = "Extensive DNA damage induced by monochloramine in gastric cells",
abstract = "Colonization of Helicobacter pylori (Hp) to gastric mucosa plays an important role for the pathogenesis of gastric mucosal lesions. We previously reported the importance of monochloramine (NH2Cl), which was derived from the interaction between Hp-urease and infiltrated leukocytes, in the course of Hp-associated gastric mucosal injury. While the long-term infection of Hp in the gastric mucosa is known to be one of the virulent factors which closely link to the gastric carcinogenesis, the details of its pathogenetic mechanisms remain speculative. The present study is designed to examine whether a NH2Cl could damage the DNA of gastric cells. Rabbit gastric mucosal cells (RGMC) or KATO III cells were cultured and suspended. Cell suspensions were exposed to HOCl, NH3 or NH2Cl for 15 min to give a final concentration of 0.1 mM. The magnitude of a double strand break of DNA was quantified by measuring the remnant double strand stained by ethidium bromide (EB), and the fluorescence intensity of EB was analyzed by spectrophotometer. Separately, cell nuclei were stained by fluorescent dye (Hoechst No. 33258) in order to evaluate the levels of chromatin condensation evoked by DNA fragmentation. The number of cells with chromatin condensation was counted. During the entire experimental period, more than 85{\%} of cells were persistently viable in all groups. NH2Cl significantly induced the DNA double strand break as well as chromatin condensation in RGMC and KATO III cells (P < 0.05). However, NH3 or HOCl did not induce the DNA double strand break as well as chromatin condensation in both cells. NH2Cl, but not its precursors (NH3 or HOCl), enhanced the levels of DNA injury, suggesting the possible involvement in the carcinogenesis of Hp-associated gastric mucosa.",
keywords = "Apoptosis, Chromatin condensation, Helicobacter pylori, Leukocyte, Urease",
author = "Hidekazu Suzuki and Mikiji Mori and Masayuki Suzuki and Kazushi Sakurai and Soichiro Miura and Hiromasa Ishii",
year = "1997",
month = "5",
day = "19",
doi = "10.1016/S0304-3835(97)04745-9",
language = "English",
volume = "115",
pages = "243--248",
journal = "Cancer Letters",
issn = "0304-3835",
publisher = "Elsevier Ireland Ltd",
number = "2",

}

TY - JOUR

T1 - Extensive DNA damage induced by monochloramine in gastric cells

AU - Suzuki, Hidekazu

AU - Mori, Mikiji

AU - Suzuki, Masayuki

AU - Sakurai, Kazushi

AU - Miura, Soichiro

AU - Ishii, Hiromasa

PY - 1997/5/19

Y1 - 1997/5/19

N2 - Colonization of Helicobacter pylori (Hp) to gastric mucosa plays an important role for the pathogenesis of gastric mucosal lesions. We previously reported the importance of monochloramine (NH2Cl), which was derived from the interaction between Hp-urease and infiltrated leukocytes, in the course of Hp-associated gastric mucosal injury. While the long-term infection of Hp in the gastric mucosa is known to be one of the virulent factors which closely link to the gastric carcinogenesis, the details of its pathogenetic mechanisms remain speculative. The present study is designed to examine whether a NH2Cl could damage the DNA of gastric cells. Rabbit gastric mucosal cells (RGMC) or KATO III cells were cultured and suspended. Cell suspensions were exposed to HOCl, NH3 or NH2Cl for 15 min to give a final concentration of 0.1 mM. The magnitude of a double strand break of DNA was quantified by measuring the remnant double strand stained by ethidium bromide (EB), and the fluorescence intensity of EB was analyzed by spectrophotometer. Separately, cell nuclei were stained by fluorescent dye (Hoechst No. 33258) in order to evaluate the levels of chromatin condensation evoked by DNA fragmentation. The number of cells with chromatin condensation was counted. During the entire experimental period, more than 85% of cells were persistently viable in all groups. NH2Cl significantly induced the DNA double strand break as well as chromatin condensation in RGMC and KATO III cells (P < 0.05). However, NH3 or HOCl did not induce the DNA double strand break as well as chromatin condensation in both cells. NH2Cl, but not its precursors (NH3 or HOCl), enhanced the levels of DNA injury, suggesting the possible involvement in the carcinogenesis of Hp-associated gastric mucosa.

AB - Colonization of Helicobacter pylori (Hp) to gastric mucosa plays an important role for the pathogenesis of gastric mucosal lesions. We previously reported the importance of monochloramine (NH2Cl), which was derived from the interaction between Hp-urease and infiltrated leukocytes, in the course of Hp-associated gastric mucosal injury. While the long-term infection of Hp in the gastric mucosa is known to be one of the virulent factors which closely link to the gastric carcinogenesis, the details of its pathogenetic mechanisms remain speculative. The present study is designed to examine whether a NH2Cl could damage the DNA of gastric cells. Rabbit gastric mucosal cells (RGMC) or KATO III cells were cultured and suspended. Cell suspensions were exposed to HOCl, NH3 or NH2Cl for 15 min to give a final concentration of 0.1 mM. The magnitude of a double strand break of DNA was quantified by measuring the remnant double strand stained by ethidium bromide (EB), and the fluorescence intensity of EB was analyzed by spectrophotometer. Separately, cell nuclei were stained by fluorescent dye (Hoechst No. 33258) in order to evaluate the levels of chromatin condensation evoked by DNA fragmentation. The number of cells with chromatin condensation was counted. During the entire experimental period, more than 85% of cells were persistently viable in all groups. NH2Cl significantly induced the DNA double strand break as well as chromatin condensation in RGMC and KATO III cells (P < 0.05). However, NH3 or HOCl did not induce the DNA double strand break as well as chromatin condensation in both cells. NH2Cl, but not its precursors (NH3 or HOCl), enhanced the levels of DNA injury, suggesting the possible involvement in the carcinogenesis of Hp-associated gastric mucosa.

KW - Apoptosis

KW - Chromatin condensation

KW - Helicobacter pylori

KW - Leukocyte

KW - Urease

UR - http://www.scopus.com/inward/record.url?scp=0030902036&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030902036&partnerID=8YFLogxK

U2 - 10.1016/S0304-3835(97)04745-9

DO - 10.1016/S0304-3835(97)04745-9

M3 - Article

C2 - 9149131

AN - SCOPUS:0030902036

VL - 115

SP - 243

EP - 248

JO - Cancer Letters

JF - Cancer Letters

SN - 0304-3835

IS - 2

ER -