Objective To investigate the effects of long non-coding ribonucleic acid (LncRNA) repulsive guidance molecules B-antisense chain (RGMB-AS1) on the proliferation, migration and invasion of human prostate cancer cells (DU145), and explore its targeting effect on miR-574-3p. Methods Human prostate cancer cells (DU145) were cultured and used in the study. Cell tests were divided into 9 groups including RGMB-AS1 upregulation group (transfected with pcDNA3. 1-RGMB-AS1), RGMB-AS1 upregulation control group (transfected with pcDNA3. 1 control), RGMB-AS1 downregulation group (transfected with si-RGMB-ASl), RGMB-AS1 downregulation control group (transfected with si-NC), miR-574-3p upregulation group (transfected with miR-574-3p mimics), miR-574-3p upregulation control group (transfected with miR mimics NC ), miR-574-3p downregulation group (transfected with miR-574-3 p inhibitor), miR-574-3p downregulated control group (transfected with miR inhibitor NC) and blank group, and 3 wells in each group. After 48 hours culture, cell proliferation activity was detected by CCK-8 assay. Cell migration ability was evaluated by wound healing assay. Cell invasive ability was assessed by Matrigel invasion assay (Transwell chamber). The expression of LncRNA RGMB-AS1, the expression of miR-574-3p, and the gene expressions of Cyclin D1, P21, ZEB1, N-cadherin, E-cadherin, and Vimentin were detected by LncRNA-RT-qPCR, miRNA-RT-qPCR and RT-qPCR, respectively. The protein expressions of Cyclin Dl, P21, ZEB1, N-cadherin, E-cadherin, and VIM were measured by western blot. The dual luciferase gene reporter assay was used to determine whether LncRNA RGMB-AS1 targets miR-574-3p. Results Compared with those in the blank group, RGMB-AS1 up-regulated control group, RGMB-AS1 down-regulated control group, miR-574-3p up-regulated control group, and miR-574-3p down-regulated control group, the D value, scratch healing rates, invasive cell counts, Cyclin Dl, ZEB1, N-cadherin, and VIM expressions in the RGMB-AS1 up-regulated group and miR-574-3p up-regulated group were all decreased (P <0. 05), while the proliferation inhibition rates, P21, and E-cadherin expressions were all increased (P <0.05). Simultaneously, the D value, scratch healing rates, invasive cell counts, Cyclin Dl, ZEB1, N-cadherin, and VIM expressions in the RGMB-AS1 down-regulated group and miR-574-3p down-regulated group were all increased (P <0. 05), while the proliferation inhibition rates, P21, and E-cadherin expressions were all decreased (P<0.05). Compared with the miR-NC group, the miR-574-3p mimics group showed a decrease in Wt-RGMR-AS1 luciferase activity (P<0. 05), but little change in Mut RGMB-AS1-luciferase activity (P > 0. 05). Conclusion Upregulation of LncRNA RGMB-AS1 can target upregulation of miR-574-3p to inhibit the proliferation, migration, and invasion of human prostate cancer cells, which may be attributed to downregulation of Cyclin D1, ZEB1, N-cadherin, VIM expressions, and upregulation of P21 and E-cadherin expressions. [ABSTRACT FROM AUTHOR]