Long non-coding RNAs (lncRNAs) have been identified to be critical mediators in various tumors associated with cancer progression. LncRNA SPRY4-IT1 serves as a novel prognostic biomarker for hepatocellular carcinoma. However, the biological role and clinical significance of lncRNA SPRY4-IT1 in human ovarian cancer (OC) need to be completely elucidated. The aim of the present study was to explore the lncRNA SPRY4-IT1 expression in human OC patients and its role in OC cells. We show that lncRNA SPRY4-IT1 expression is significantly upregulated in ovarian tumor tissues and OC cell lines in comparison with adjacent non-tumor control tissues and the human ovarian immortalized nontumorigenic ovarian surface epithelial (IOSE), respectively. Further analysis by Kaplan-Meier survival analysis and multivariate analysis indicated that high lncRNA SPRY4-IT1 expression may be an independent prognostic factor for progression-free survival (PFS) and overall survival (OS) in OC patients. Furthermore, the area under the receiver operating characteristic (ROC) curve of lncRNA SPRY4-IT1 was up to 0.8512, indicating lncRNA SPRY4-IT1 has diagnostic values to discriminate tumor tissues from nontumorous tissues. Also, knockdown of lncRNA SPRY4-IT1 inhibited the proliferation of OC cells by CCK-8 assay and clonogenic assay and arrested cell cycle at a G0/G1 stage in OC cells. In conclusion, these results suggest that lncRNA SPRY4-IT1 may be considered as a new predictor in the clinical prognosis of OC patients.