The study investigated whether intakes of fat and fatty acids are associated with the development of gastric cancer. A case-control study was conducted in the Clinical Centre Nis (Serbia) from 2005 to 2006. A total of 102 patients with histologically confirmed gastric cancer were compared with 204 controls chosen among the subjects admitted to the same hospital as cases of acute non-neoplastic diseases. A food-frequency questionnaire (FFQ) was used to assess information on the frequency of consumption of foods in the course of one year prior to cancer diagnosis or hospital admission (for controls). Nutrient values in food were obtained from the Food composition table. We did not observe statistically significant association in frequency of consumption of visible fat (sunflower oil, olive oil, lard, margarine butter) between cases and controls (p>0.05). According to the univariate logistic regression analysis, total fat (OR for highest tertile 0.80, 95%CI: 0.28-2.25), saturated fatty acids (0.85, 95%CI: 0.31-2.35), monounsaturated fatty acids (0.57, 95%CI: 0.22-1.48) and cholesterol (0.79; 95%CI:0.37-2.28) intake were not associated, and polyunsaturated fatty acids were inversely associated with the risk of gastric cancer (0.32; 95%CI: 0.12-0.83). Our results suggest that dietary polyunsaturated fat intake may help to prevent gastric cancer. Further studies are needed to confirm these results.