AbstractUse of artificial sweeteners (AS) such as aspartame, cyclamate, saccharin and sucralose is widespread. We evaluated the association of use of aspartame and other AS with cancer. In total 1881 colorectal, 1510 breast, 972 prostate and 351 stomach cancer and 109 chronic lymphocytic leukaemia (CLL) cases and 3629 population controls from the Spanish Multicase‐Control (MCC‐Spain) study were recruited (2008‐2013). The consumption of AS, from table‐top sweeteners and artificially sweetened beverages, was assessed through a self‐administered and validated food frequency questionnaire (FFQ). Sex‐specific quartiles among controls were determined to compare moderate consumers (<third quartile) and high consumers (≥ third quartile) vs non consumers (reference category), distinguishing aspartame‐containing products and other AS. Unconditional logistic regression models were used to estimate adjusted OR and 95%CI, and results were stratified by diabetes status. Overall, we found no associations between the consumption of aspartame or other AS and cancer. Among participants with diabetes, high consumption of other AS was associated with colorectal cancer (OR = 1.58, 95% CI 1.05‐2.41, P trend = .03) and stomach cancer (OR = 2.27 [0.99‐5.44], P trend = .06). High consumption of aspartame, was associated with stomach cancer (OR = 2.04 [0.7‐5.4], P trend = .05), while a lower risk was observed for breast cancer (OR = 0.28 [0.08‐0.83], P trend = .03). In some cancers, the number of cases in participants with diabetes were small and results should be interpreted cautiously. We did not find associations between use of AS and cancer, but found associations between high consumption of aspartame and other AS and different cancer types among participants with diabetes.