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基金项目:"天山英才"医药卫生高层次人才培养项目（TSYC202301B078）；省部共建中亚高发病成因与防治国家重点实验室开放课题（SKL-HIDCA-2024-KY4）；新疆维吾尔自治区自然科学基金-面上项目（2021D01C017）；新疆人工智能影像辅助诊断重点实验室开放课题（XJRGZN2024011）

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中文摘要:

摘要目的：探究miR-489-3p在前列腺癌中的作用，以及与TWIST相关蛋白1（TWIST1）的相关性。方法：选择2022年2月至2023年8月喀什地区第一人民医院泌尿外科收治的前列腺癌患者共26例，分别收集患者的前列腺癌组织和相应的癌旁正常组织。采用CancerMIRNome软件分析miR-489-3p在前列腺癌组织中的表达水平，采用GEPIA2软件分析TWIST1与前列腺癌症患者生存期关系，采用TargetScanHuman软件和双荧光素酶报告基因实验分析miR-489-3p与TWIST1靶向关系和结合位点预测。将人前列腺癌细胞PC-3分为对照组、mimics NC组、miR-489-3p mimics组、miR-489-3p mimics+vector组和miR-489-3p mimics+TWIST1组。采用MTT实验和克隆形成实验检测PC-3细胞增殖，流式细胞术检测PC-3细胞凋亡能力，细胞划痕愈合实验检测PC-3细胞迁移能力，Transwell实验检测PC-3细胞侵袭能力。采用qRT-PCR分析组织和细胞中miR-489-3p和TWIST1基因表达水平，Western blot检测TWIST1、E-cadherin、N-cadherin和Vimentin蛋白表达水平。结果：与正常组织比较，前列腺癌组织中miR-489-3p表达水平降低（P<0.05），TWIST1的mRNA和蛋白表达水平均升高（P<0.05），且TWIST1高表达患者的生存期低于TWIST1低表达患者（P<0.05）。TargetScanHuman软件在线分析和双荧光素酶报告基因实验结果显示，miR-489-3p与TWIST1存在靶向结合位点。与对照组或mimics NC组比较，miR-489-3p mimics组细胞中miR-489-3p表达水平和E-cadherin蛋白表达水平均升高（P<0.05），TWIST1 mRNA以及TWIST1、N-cadherin和Vimentin蛋白表达水平均降低（P<0.05），细胞克隆形成数、增殖活性、细胞相对迁移率和侵袭细胞数量均降低（P<0.05），细胞凋亡率升高（P<0.05）。与miR-489-3p mimics+vector组比较，miR-489-3p mimics+TWIST1组细胞中miR-489-3p表达水平和E-cadherin蛋白表达水平均降低（P<0.05），TWIST1 mRNA以及TWIST1、N-cadherin和Vimentin蛋白表达水平均升高（P<0.05），细胞克隆形成数、增殖活性、细胞相对迁移率和侵袭细胞数量均升高（P<0.05），细胞凋亡率降低（P<0.05）。结论：miR-489-3p可能是前列腺癌发展的抑制因子，过表达miR-489-3p可以促进PC-3细胞凋亡，抑制PC-3细胞增殖、迁移、侵袭和上皮间质转化过程，该作用可能与抑制TWIST1表达有关。

英文摘要:

ABSTRACT 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 expression 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 decreased(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.

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