Many reproductive factors are associated with breast cancer risk, potentially through a hormonal pathway. The peptide hormone prolactin is essential in mammary development and lactation and may be a link between risk factors and breast cancer. While higher prolactin levels are associated with increased breast cancer risk, few determinants of prolactin levels are known. We conducted a cross-sectional analysis among 1,089 premenopausal and 1,311 postmenopausal women within the Nurses' Health Study (NHS) and the NHS II to examine the associations of reproductive factors, benign breast disease and family history of breast cancer with plasma prolactin levels. Parous women had significantly lower prolactin levels than nulliparous women (parous vs. nulliparous multivariate-adjusted geometric means 5 14.1 ng/mL vs. 16.6 ng/mL, p < 0.001 for premenopausal and 9.1 vs. 10.1, p 5 0.04 for postmenopausal women), although levels did not decrease with increasing number of children for either premenopausal (p-trend 5 0.23) or postmenopausal (p-trend 5 0.07) parous women. Age at first birth was not associated with prolactin levels. The reduction in prolactin levels among parous premenopausal women appeared to attenuate with increasing time since first birth, but the trend was not statistically significant (p-trend 5 0.12). Age at menarche, duration of lactation and benign breast disease were not associated with prolactin levels. Family history of breast cancer was associated with significantly higher prolactin levels when compared with no family history among premenopausal (15.9 ng/mL vs. 14.3 ng/ mL, p 5 0.04) but not postmenopausal (p 5 0.73) women. In conclusion, the associations of parity and family history with breast cancer risk may be mediated, at least in part, by prolactin levels. ' 2006 Wiley-Liss, Inc.Key words: prolactin; reproductive factors; family history Many reproductive factors consistently associated with breast cancer risk, such as age at menarche, parity and age at first birth, likely affect risk through a hormonal pathway. 1 The peptide hormone prolactin, essential in mammary development and lactation, is associated with an increased risk of breast cancer in both premenopausal 2 (Tworoger et al., 2006, submitted) and postmenopausal 3-6 women, likely through an increase in cell proliferation and inhibition of apoptosis. 7 Thus, prolactin may be one of potentially many hormonal links between reproductive risk factors and the incidence of breast cancer. 7,8 While the association between prolactin levels and breast cancer risk is fairly consistent, whether lifestyle or reproductive factors influence prolactin levels is not well known. A number of cross-sectional studies have observed an inverse association between parity and prolactin levels in both premenopausal 9-13 and postmenopausal 9,14 women, although whether prolactin levels decline further after the first pregnancy is unclear. Other birthrelated factors, such as age at first birth, time since last birth and history of lactation, have been assessed in ...