Department of Urology, the Second Affiliated Hospital of Xi'an Jiaotong University.
Department of Nursing, Xi'an Beifang Chinese Medicine Skin Disease Hospital, Xi'an, Shaanxi, China.
Anticancer Drugs. 2020 Jul;31(6):592-600. doi: 10.1097/CAD.0000000000000882.
Our study aimed to further investigate the roles and molecular mechanisms of lncRNA taurine upregulated gene 1 (TUG1) in the development and progression of PC. RT-qPCR assay was carried out to measure expression of TUG1, miR-496, together with β-catenin, cyclin D1 and c-myc. Protein levels of β-catenin, cyclin D1 and c-myc were detected by western blot assay. Cell proliferative ability was assessed by colony formation assay and CCK-8 assay. Cell migratory and invasive capacities were evaluated by Transwell migration and invasion assay. The interaction between miR-496 and TUG1 was explored by bioinformatics analysis, luciferase reporter assay and RNA immunoprecipitation assay. Mouse xenograft experiments were performed to further investigate the roles and molecular basis of TUG1 in the tumorigenesis of PC in vivo. TUG1 was highly expressed in PC tissues and cells (DU145 and PC3). TUG1 knockdown inhibited proliferation, migration and invasion in DU145 and PC3 cells. Moreover, TUG1 suppressed miR-496 expression by direct interaction. TUG1 overexpression abrogated miR-96-mediated antiproliferation, anti-migration and anti-invasion effects in DU145 and PC3 cells. TUG1 knockdown inactivated Wnt/β-catenin signaling pathway by upregulating miR-496 in DU145 and PC3 cells. Additionally, TUG1 knockdown inhibited DU145 cells derived PC xenograft growth by upregulating miR-496 and inactivating Wnt/β-catenin signaling in vivo. TUG1 knockdown suppressed PC cell proliferation, migration and invasion in vitro and curbed PC xenograft growth in vivo by regulating miR-496/Wnt/ β-catenin signaling pathway, deepening our understanding on etiology of PC.
我们的研究旨在进一步探讨长链非编码 RNA 牛磺酸上调基因 1（TUG1）在前列腺癌（PC）发展和进展中的作用和分子机制。通过实时定量 PCR 检测 TUG1、miR-496 以及β-连环蛋白、细胞周期蛋白 D1 和 c-myc 的表达。通过 Western blot 检测β-连环蛋白、细胞周期蛋白 D1 和 c-myc 的蛋白水平。通过集落形成实验和 CCK-8 实验评估细胞增殖能力。通过 Transwell 迁移和侵袭实验评估细胞迁移和侵袭能力。通过生物信息学分析、荧光素酶报告基因检测和 RNA 免疫沉淀实验探索 miR-496 与 TUG1 之间的相互作用。通过小鼠异种移植实验进一步研究 TUG1 在体内致前列腺癌中的作用和分子基础。TUG1 在前列腺癌组织和细胞（DU145 和 PC3）中高表达。TUG1 敲低抑制 DU145 和 PC3 细胞的增殖、迁移和侵袭。此外，TUG1 通过直接相互作用抑制 miR-496 的表达。TUG1 过表达可消除 miR-96 介导的对 DU145 和 PC3 细胞的增殖、迁移和侵袭的抑制作用。TUG1 敲低通过上调 miR-496 在 DU145 和 PC3 细胞中使 Wnt/β-连环蛋白信号通路失活。此外，TUG1 敲低通过体内上调 miR-496 和失活 Wnt/β-连环蛋白信号通路抑制 DU145 细胞衍生的前列腺癌异种移植物的生长。TUG1 敲低通过调节 miR-496/Wnt/β-连环蛋白信号通路抑制体外 PC 细胞的增殖、迁移和侵袭，并抑制体内前列腺癌异种移植物的生长，从而加深我们对前列腺癌病因的理解。
