[Abstract] Objective: To investigate the expression changes of miR-96 in endometrial cancer tissues and cells, and to explore its effect on tumor malignant phenotypes as well as the possible mechanisms. Methods:From April 2016 to December 2018, 76 cases of endometrial cancer tissues from 76 patients who were surgically treated in the Department of Obstetrics and Gynecology of our hospital were selected for this study. qPCR was used to detect the expression of miR-96 in human endometrial cancer tissues and cells, and the correlation between the miR-96 expression and the clinicopathological characteristics of patients was analyzed. miR-96 inhibitor was transfected into human endometrial cancer Ishikawa cells in vitro. After transfection, the expression of miR-96 in Ishikawa cells was detected by qPCR; the tumor biological behaviors of Ishikawa cells were detected by CCK-8 test, Clone formation test, Flow cytometry, Scratch test and Transwell test; and the FOXO1 protein expression in Ishikawa cells was detected by WB. At the same time, Dual luciferase reporter gene assay was used to observe the targeting relationship between miR-96 and FOXO1. Results: The results of qPCR showed that the expression of miR-96 was abnormally high in human endometrial cancer cells (JEC, Ishikawa, HEC-1B) and endometrial cancer tissues (all P<0.01), and the expression of miR-96 was closely related to FIGO stage and lymph node metastasis (all P<0.05). After transfection with miR-96 inhibitor, the expression level of miR-96 in Ishikawa cells decreased significantly (P<0.01), the proliferation activity and clone formation ability decreased significantly (all P<0.01), the apoptotic rate increased significantly (P<0.01), and the scratch healing rate and the number of invasive transmembrane cells decreased significantly (P<0.01). Dual luciferase reporter gene assay showed that miR-96 could directly target FOXO1, and WB showed that miR-96 could negatively regulate FOXO1 protein expression in Ishikawa cells (P<0.01). Conclusion: The expression of miR-96 is abnormally high in endometrial cancer tissues and cells. Inhibiting the expression of miR-96 can inhibit the proliferation, invasion and migration of endometrial cancer cells and promote their apoptosis. The mechanism may be related to the targeted regulation of FOXO1.