LncRNA RGMB-AS1 targeting miR-574-3p affects the proliferation,migration and invasion of human prostate cancer cells(DU145)
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-AS1),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-574-3p 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 D1,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 D1,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 D1,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-RGMB-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.
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