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Am J Physiol Gastrointest Liver Physiol 279: G1169-G1176, 2000;
0193-1857/00 $5.00
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Vol. 279, Issue 6, G1169-G1176, December 2000

Regulation of growth and apoptosis of cultured guinea pig gastric mucosal cells by mitogenic oxidase 1

Shigetada Teshima1, Hiromu Kutsumi2, Tsukasa Kawahara1, Kyoichi Kishi1, and Kazuhito Rokutan1

1 Department of Nutrition, School of Medicine, The University of Tokushima, Tokushima City, Tokushima 770-8503; and 2 Gastrointestinal Unit, Kyoto Red Cross Hospital, Kyoto City, Kyoto 605-0981, Japan

We previously reported that primary cultures of guinea pig gastric pit cells expressed all of the phagocyte NADPH oxidase components (gp91-, p22-, p67-, p47-, and p40-phox) and could spontaneously release superoxide anion (O2-). We demonstrate here that pit cells express a nonphagocyte-specific gp91-phox homolog (Mox1) but not gp91-phox. Inclusion of catalase significantly inhibited [3H]thymidine uptake during the initial 2 days of culture. Pit cells, matured on day 2, slowly underwent spontaneous apoptosis. Scavenging O2- and related oxidants by superoxide dismutase plus catalase or N-acetyl cysteine (NAC) and inhibiting Mox1 oxidase by diphenylene iodonium activated caspase 3-like proteases and markedly enhanced chromatin condensation and DNA fragmentation. This accelerated apoptosis was completely blocked by a caspase inhibitor, z-Val-Ala-Asp-CH2F. Mox1-derived reactive oxygen intermediates constitutively activated nuclear factor-kappa B, and inhibition of this activity by nuclear factor-kappa B decoy oligodeoxynucleotide accelerated their spontaneous apoptosis. These results suggest that O2- produced by the pit cell Mox1 oxidase may play a crucial role in the regulation of their spontaneous apoptosis as well as cell proliferation.

NADPH oxidase; superoxide anion; hydrogen peroxide; antiapoptosis; cell growth


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