Objective To investigate the effect of HOX transcriptional antisense RNA (HOTAIR) targeted regulating miR-424 expression on the biological behavior of ovarian cancer cells. Methods Sixty ovarian cancer samples from patients with ovarian cancer and randomly matched 60 benign ovarian tumor tissues were collected during January to June 2018 in the Third Affiliated Hospital of Zunyi Medical University. q-PCR was used to detect the expression of HOX transcript antisense RNA (HOTAIR) in ovarian cancer and adjacent tissues, and analyze the survival rate of ovarian cancer patients with different HOTAIR expression levels, analyze the relationship between the expression level of HOTAIR and the clinicopathological parameters of ovarian cancer patients. A2780 ovarian cancer cells were transfected with small interfering RNA (siRNA) to construct siRNA-HOTAIR (si-HOTAIR) cells as the experimental group, and A2780 ovarian cancer cells transfected with siRNA-NC (si-NC) cells were used as the control group. Dual fluorescence was used to detect the interaction between HOTAIR and miR-424 by the enzyme reporter gene,A2780 cells were used in scratch test to detect the change in the migration ability of ovarian cancer cells after inhibiting HOTAIR,Transwell invasion test was used to detect the change in the invasive ability of ovarian cancer cells after inhibiting HOTAIR, and Flow cytometry was performed to analyze the changes of ovarian cancer cell apoptosis after inhibiting HOTAIR. The nude mouse subcutaneous tumor formation experiment was used to detect the effect of inhibiting HOTAIR on the tumor size and volume of ovarian cancer cells. Results qPCR results showed that the expression of HOTAIR increased significantly in ovarian cancer tissues than in benign ovarian tumor tissues [(58.64±6.32) vs. (6.35±0.02), P<0.05]. The long term survival rate was lower in patients with high expression level of HOTAIR than in patients with low expression level of HOTAIR. Dual-Luciferase reporter gene system showed that HOTAIR can specifically bind to the 3'-UTR of miR-424 [(0.42±0.08) vs. (1.06±0.11), P<0.05]. Scratch test and Transwell invasion test showed that, compared with control group, inhibiting the expression of HOTAIR can inhibit the migration behavior [(21.21±3.08) μm vs. (92.34±8.65) μm, P<0.05] and invasion behavior of ovarian cancer cells in experimental group(28.63%±5.59% vs. 275.36%±21.62%, P<0.05). The results of flow cytometry showed that inhibiting the expression of HOTAIR can enhance the apoptosis of ovarian cancer cells in experimental group (30.2%±2.12% vs. 7.31%±2.01%, t=10.64, P<0.05). In vivo tumor formation experiments in nude mice showed that, compared with control group, the tumor volume and weight of tumor-bearing mice correspondingly decreased [(0.85±0.06) cm³ vs. (3.05±0.28) cm³, t=13.41, P<0.05; (1.12±0.08) g vs. (2.91±0.19) g,t=11.64, P<0.05] after inhibiting HOTAIR expression. Conclusion lncRNA HOTAIR can target miR-424 to regulate the apoptosis, migration and invasion of ovarian cancer cells.