Dietary factors play essential roles in gastric carcinogenesis. We recently found that dietary supplementation with NaHCO 3 significantly increased the development of gastric cancer in a rat gastric stump model. Here, we analysed nontransformed gastric mucosa for expression of the cancer-related proteins cyclooxygenase-2 (COX-2) and ornithine decarboxylase (ODC), and we examined the relationship between expression levels of those proteins and mucosal proliferation. Research has shown that COX-2 is upregulated in gastric mucosal inflammation and is strongly associated with gastrointestinal cancer. ODC is the key enzyme in polyamine synthesis and a regulator of cell proliferation. We performed gastric resections on 48 Wistar rats to induce spontaneous gastric cancer; half of these animals were given a normal diet, and the other half received a diet supplemented with NaHCO 3 . Twentyfour unoperated rats served as a control group. The surgical procedure per se led to a significant rise in mucosal expression of COX-2 and an associated increase in cell proliferation. However, the COX-2 level in gastric mucosa was not further affected by dietary supplementation of carbonate. Interestingly, nontransformed gastric mucosa in the operated rats receiving a carbonate-supplemented diet showed a pronounced increase in ODC expression that was strongly correlated with a further enhanced cell proliferation. These results indicate that carbonate ions, which represent a major constituent of intestinal reflux into the stomach, increase the expression of ODC and thereby enhance cell proliferation in nontransformed mucosa, and consequently elevate the risk of gastric cancer. ' 2007 Wiley-Liss, Inc.Key words: experimental gastric cancer; inflammation; carbonate ions; ornithine decarboxylase; proliferation Gastric adenocarcinoma is the second leading cause of cancer death worldwide.1 Prognosis is poor in advanced disease, with a 5 year overall survival of only 7-20%, 2,3 and thus detecting stomach cancer in its early stages is pivotal for a good outcome. However, disappointingly, this is done in only 10-20% of the patients in many Western countries, such as the United States, although the proportion of detections is as high as 50% in some nations, including Japan.4 Therefore, prevention of gastric cancer has become a principal objective, and, during the last 5-10 years, studies with that goal have focused on the role of H. pylori infections and diet. 5,6 The importance of dietary supplements in relation to an elevated risk of gastric cancer is underlined by the high incidence of this disease in Japan, 1 where the diet is generally high in nitrosamines, salt, and smoked or cured food, which are known to increase the risk of developing gastric adenocarcinoma. 7,8 However, reliable quantitative data are lacking with regard to how various ions in the diet can affect the incidence of gastric cancer.9,10 We recently investigated calcium as a dietary supplement, considering its role in either protecting against or elevating the risk of gastric...