Introduction: Estrogen, both endogenous and exogenous, is generally recognized as a potential risk factor in endometrial cancer. It is also known that postmenopausal estrogen use is involved in endometrial cancer pathophysiology. In this case control study, we analyzed whether having gallstones or a cholecystectomy, which are much more common in obese women or those who use exogenous estrogen, might be associated with endometrial cancer risk. Material, methods: A total of 470 patients with endometrial cancer were enrolled in the study along with 369 control women of comparable age without history of malignancy. Participants completed a structured questionnaire focused mainly on the reproductive history of the patients, history of cholecystectomy or gallstones and postmenopausal replacement therapy. Information on tumor staging or grading were obtained from the medical records along with cholesterol plasma levels. BMI at the time of diagnosis was calculated. Results: No case control differences in cholecystectomy frequency were observed between the endometrial cancer patients and controls. Endometrial cancer was found to be associated with obesity, age and low parity. Age of menarche was significantly associated to estrogen receptor presence in tumor mass (p=0.03), Discussion: As lack of association was observed, it might be concluded that cholecystectomy or gallstones do not represent a significant risk factor for endometrial cancer development in the Czech population.