Irregular working hours, including working at night, have serious psychological and physiological effects. In a nationwide population-based case-control study, we investigated the breast cancer risk among 30- to 54-year-old Danish women who worked predominantly at night. Individual employment histories were reconstructed back to 1964 for each of 7035 women with breast cancer and their individually matched controls from the records of a nationwide pension scheme with compulsory membership. Odds ratios, including 5 years of induction time and adjusted for socio-economic status, age at the birth of first and last child and number of children, were estimated by conditional logistic regression analysis. The odds ratio for breast cancer among women who worked at night at least half of a year was 1.5 (95% confidence interval, 1.2 to 1.7), and there was a tendency to increasing odds ratio by increasing duration of nighttime employment.