To examine the hypothesis that tobacco smoke is associated with the risk of female breast cancer, we estimated the relative risks of active and passive smoke in middle-aged Japanese women in a population-based prospective study. The cohort consisted of residents in 4 public health center areas, aged 40 to 59 years. A self-administered questionnaire survey was conducted in 1990. This analysis included 21,805 subjects, 180 of whom had developed breast cancer by December 31, 1999. When the reference was defined as never-active smokers without passive smoking, adjusted relative risks (RRs) were 1.9 (95% confidence interval [CI] ؍ 1.0 -3.6) in current active smokers, 1.2 (95% CI ؍ 0.4 -4.0) in ex-active smokers and 1.2 (95% CI ؍ 0.8 -1.6) in never-active smokers with passive smoking. The elevated risk for ever-smokers was clearly observed in premenopausal women at baseline (RR ؍ 3.9, 95% CI ؍ 1.5-9.9) but not in postmenopausal women (RR ؍ 1.1, 95% CI ؍ 0.5-2.5). In never-active smokers, the adjusted RR for passive smoking, residential or occupational/public tobacco smoke exposure was 1.1 (95% CI ؍ 0.8 -1.6). In premenopausal women, passive smoking increased the risk (RR ؍ 2.6; 95% CI ؍ 1.3-5.2) but not in postmenopausal women (RR ؍ 0.7; 95% CI ؍ 0.4 -1.0). We conclude that tobacco smoking increases the risk of female breast cancer in premenopausal women.