Women enrolled in the Nurses’ Health Study provided the data for this prospective, nested, case-control study. A total of 79,439 women, with no history of cardiovascular disease or cancer, were followed-up for 24 years to determine the relationship between aspirin use and mortality risk. The relative risk of all-cause mortality was reduced in women who reported current aspirin use compared to those with no regular aspirin use (RR 0.75; 95% CI 0.71-0.81). Mortality from cardiovascular disease (RR 0.62; 95% CI 0.55-0.71) and cancer (RR 0.88; 95% CI 0.81-0.96) was also reduced. Cardiovascular mortality benefits were achieved with 1-5 years of aspirin use (RR 0.75; 95% CI 0.61-0.92), whereas risk of cancer mortality was not significantly reduced until after 10 years of aspirin use (Plinear trend = 0.005). Only low and moderate aspirin doses were beneficial. Greater benefits were observed in older participants and those with more cardiovascular risk factors.