miR-489-3p Inhibits the Malignant Development of Prostate Cancer by Targeting TWIST1
Objective:To investigate the role of miR-489-3p in prostate cancer and its correlation with TWIST-related protein 1(TWIST1).Methods:A total of 26 patients with prostate cancer admitted to the department of urology of the First People's Hospital of Kashi from February 2022 to August 2023 were selected,and prostate cancer tissues and corresponding adjacent normal tissues of patients were collected,respectively.The expression level of miR-489-3p in prostate cancer tissues was analyzed by CancerMIRNome software,and the relationship between TWIST1 and survival of prostate cancer patients was analyzed by GEPIA2 software.The targeting relationship between miR-489-3p and TWIST1 and its binding site was analyzed by TargetScanHuman software and double luciferase reporter gene assay.Human prostate cancer cells PC-3 were divided into control group,mimics NC group,miR-489-3p mimics group,miR-489-3p mimics+vector group and miR-489-3p mimics+TWIST1 group.The proliferation of PC-3 cell was detected by MTT assay and colony formation assay,apoptosis of PC-3 cell was detected by flow cytometry,migration of PC-3 cell was detected by scratch healing experiment,and invasion of PC-3 cell was detected by Transwell assay.The gene expression levels of miR-489-3p and TWIST1 in tissues and cells were analyzed by qRT-PCR.The protein expression levels of TWIST1,E-cadherin,N-cadherin and Vimentin were detected by Western blot.Results:Compared with normal tissues,the expression level of miR-489-3p in prostate cancer tissues was decreased(P＜0.05),while mRNA and protein expression levels of TWIST1 were increased(P＜0.05),and the survival time of patients with high expres-sion of TWIST1 was lower than that of patients with low expression of TWIST1(P＜0.05).The results of TargetScanHuman analysis and double luciferase reporter gene experiment showed that miR-489-3p had a targeted binding site with TWIST1.Compared with control group or the mimics NC group,the expression level of miR-489-3p and E-cadherin protein in miR-489-3p mimics group were increased(P＜0.05).The expression levels of TWIST1 mRNA,and the protein expression levels of TWIST1,N-cadherin and Vimentin were de-creased(P＜0.05).The number of cell clone formation,proliferation activity,cell relative mobility and the number of invasive cells were decreased(P＜0.05),and the apoptosis rate was increased(P＜0.05).Compared with miR-489-3p mimics+vector group,the expression level of miR-489-3p and E-cadherin protein in miR-489-3p mimics+TWIST1 group were decreased(P＜0.05).The expression levels of TWIST1 mRNA,and the protein expression levels of TWIST1,N-cadherin and Vimentin were increased(P＜0.05).The number of cell clone formation,proliferation activity,cell relative mobility and the number of invasive cells were increased(P＜0.05),and the apoptosis rate was decreased(P＜0.05).Conclusion:miR-489-3p is an inhibitor of the development of prostate cancer.Overexpression of miR-489-3p can promote apoptosis of PC-3 cells,and inhibit the proliferation,migration,invasion and EMT process of PC-3 cells,which may be related to the inhibition of the expression level of TWIST1.
miR-489-3pTWIST1Prostate cancerProliferation and apoptosisMigration and invasionEpithelial-mesenchymal transition(EMT)
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