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Am J Physiol Gastrointest Liver Physiol 275: G712-G716, 1998;
0193-1857/98 $5.00
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Vol. 275, Issue 4, G712-G716, October 1998

Monochloramine induced DNA fragmentation in gastric cell line MKN45

Hidekazu Suzuki1, Koichi Seto2, Mikiji Mori1, Masayuki Suzuki3, Soichiro Miura4, and Hiromasa Ishii1

1 Department of Internal Medicine, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582; 2 Department of Pharmacology, Central Research Laboratory, Zeria Pharmaceutical Co., Ltd., Saitama 369-0100; 3 Department of Gastroenterology, National Tokyo Medical Center, Meguro-ku, Tokyo 152-0021; and 4 Second Department of Internal Medicine, National Defense Medical College, Saitama 359-8513, Japan

Monochloramine (NH2Cl) is known to be one of the virulence factors in Helicobacter pylori-associated gastric mucosal injury. The present study was designed to examine NH2Cl-evoked DNA fragmentation in the gastric epithelial cell line MKN45. NH2Cl was produced by mixing NH3 with sodium hypochlorite (NaClO). MKN45 cells were exposed to NH2Cl, NH3, or NaClO in Hanks' balanced salt solution. DNA cleavage was evaluated quantitatively by photometeric enzyme immunoassay for the in vitro determination of cytoplasmic mono- and oligonucleosomes. Damage to the plasma membrane was assessed by measuring the activity of lactate dehydrogenase in the supernatants. Separately, DNA ladder formation was performed to confirm the incidence of DNA fragmentation. NH2Cl (0.001-0.01 mM) significantly increased the cytoplasmic mono- and oligonucleosomes, suggesting the incidence of DNA cleavage. The DNA ladder was clearly evoked by NH2Cl. NH2Cl induced a DNA fragmentation, one of the important aspects in apoptosis, in the gastric cell line MKN45.

Helicobacter pylori; leukocyte; mononucleosome; oligonucleosome; lactate dehydrogenase


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