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OBJECTIVE The aim of this study was to investigate the potential effect of microRNA-135b-5p (miR-135b-5p) on the development of ovarian cancer (OC) and to explore the relevant mechanism. PATIENTS AND METHODS The expression of miR-135b-5p in OC tissues and cells was detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). MicroRNA online prediction websites were used to screen the potential targets of miR-135b-5p. Subsequently, luciferase reporter gene assay and Western blot (WB) were performed for further confirmation. In addition, the effects of miR-135b-5p on cell function were analyzed by relevant experiments in vitro. RESULTS MiR-135b-5p was lowly expressed in both OC tissues and cell lines. Combined with online prediction software, luciferase reporter gene assay and WB, KDM5B was predicted and verified as a downstream target gene of miR-135b-5p. Down-regulating the expression of KDM5B by over-expressing miR-135b-5p in OC cells could effectively control the proliferation and apoptosis of OC cells. Cell proliferation was significantly reduced, while cell apoptosis was promoted after miR-135b-5p mimics transfection in OC cells. CONCLUSIONS By targeting KDM5B, miR-135b-5p exerted an excellent anti-cancer effect in OC cells. Our findings indicated that miR-135b-5p/KDM5B might become a feasible and new target of OC treatment.