Bortezomib suppresses ubiquitin (Ub)-dependent protein degradation and preferentially kills various tumour cells in vitro and in animal models. However, its mechanism of action is not fully understood. We report that bortezomib inhibits the proliferation and proteasomal activity of human endometrial cancer cells and induces G2/M arrest and apoptosis by modulating the miRNA level. By miRNA microarray, iR-17-5p was the most downregulated of all those in HTB-111 and Ishikawa cells after bortezomib treatment. This observation was confirmed by quantitative real-time PCR (qRT-PCR). Target prediction using TargetScan software identified p21 as a potential target for miR-17-5p, which was confirmed by luciferase reporter, qRT-PCR and Western blot assays. The transfection of miR-17-5p mimics or siRNA-p21 reversed the effect of bortezomib on HTB-111 and Ishikawa cells, indicating that miR-17-5p may mediate the function of bortezomib by targeting p21 in endometrial cancer cells. These findings show novel mechanisms by which bortezomib inhibits proliferation and promotes the apoptosis of human endometrial cancer cells.