To investigate the relationship between trans fatty acids and postmenopausal breast cancer in European populations differing greatly in their dietary fat intakes, a case control study using adipose tissue stores of trans fatty acids as a biomarker of exposure was conducted. Subjects included 698 postmenopausal incident cases of primary breast cancer and controls randomly drawn from local population and patient registries, ages 50-74 Concentrations of individual trans fatty acids in gluteal fat biopsies were measured in these women. The adipose concentration of trans fatty acids showed a positive association with breast cancer. The covariate-adjusted association with breast cancer. The covariate-adjusted OR was 1.40 (95% confidence interval: 1.02, 1.93) for the difference between the 75th and 25th percentiles of total adipose trans. The adjusted OR for trans in the lowest tertile of polyunsaturated fatty acid reached 3.6 (2.2, 6.1). These associations were not attributable to differences in age, body mass index, exogenous hormone use, or socioeconomic status. These findings suggest an association of adipose stores of trans fatty acids with postmenopausal breast cancer in European women. They require confirmation in other populations, with concomitant consideration of the potential roles of dietary saturated and monounsaturated fats.