The Lncrna-TUG1/EZH2 Axis Promotes Pancreatic Cancer Cell Proliferation, Migration and EMT Phenotype Formation Through Sponging Mir-382
The Lncrna-TUG1/EZH2 Axis Promotes Pancreatic Cancer Cell Proliferation, Migration and EMT Phenotype Formation Through Sponging Mir-382
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DOI：
10.1159/000479990
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Zhou, Mengtao
中科院分区：
文献类型：
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作者：
Zhao, Liang;Sun, Hongwei;Zhou, Mengtao
背景/目的：胰腺癌（Pancreatic carcinoma，PC）是世界上最常见的恶性肿瘤之一。牛磺酸上调基因1（taurine upregulated gene 1，TUG 1）是一种被牛磺酸上调的转录本，在多种肿瘤中均有异常表达的报道。然而，TUG 1在PC中的生物学作用和分子机制仍需进一步研究。方法：采用实时荧光定量PCR（qRT-PCR）方法检测TUG 1在PC细胞系和组织中的表达。MTT法和集落形成实验检测TUG 1对细胞增殖的影响。采用创伤愈合实验、transwell实验和western blot实验检测TUG 1对细胞迁移和上皮间质转化（EMT）表型的影响。通过RNA结合蛋白免疫沉淀（RIP）和生物素-亲和素下拉系统（biotin-avidin pulldown system）证实miR-328和TUG 1之间的相互作用。使用临床样品和RT-qPCR的基因表达阵列分析表明，zeste同源物2（EZH 2）的增强子是PC中miR-382的靶标。结果：在本研究中，我们报道了TUG 1在PC组织和细胞系中过表达，并且TUG 1的高表达预示着预后不良。进一步的实验表明，过表达的TUG 1促进细胞增殖，迁移，并有助于EMT的形成，而沉默的TUG 1导致相反的结果。此外，荧光素酶报告基因测定、RIP测定和RNA下拉测定证明TUG 1可以竞争性地海绵化miR-382，从而调节EZH 2。结论：总的来说，这些发现揭示了TUG 1作为致癌IncRNA发挥作用，其至少部分地通过作为内源性“海绵”发挥作用并竞争miR-382与miRNA靶标EZH 2的结合来促进肿瘤进展。(C)2017作者（s）由S. Karger AG，巴塞尔
Background/Aims: Pancreatic carcinoma (PC) is the one of the most common and malignant cancers worldwide. LncRNA taurine upregulated gene 1 (TUG1) was initially identified as a transcript upregulated by taurine, and the abnormal expression of TUG1 has been reported in many cancers. However, the biological role and molecular mechanism of TUG1 in PC still needs further investigation. Methods: Quantitative real-time PCR (qRT-PCR) was performed to measure the expression of TUG1 in PC cell lines and tissues. MTT and colony formation assays were used to measure the effect of TUG1 on cell proliferation. A wound healing assay, transwell assay and western blot assay were employed to determine the effect of TUG1 on cell migration and the epithelial mesenchymal transition (EMT) phenotype. RNA-binding protein immunoprecipitation (RIP) and a biotin-avidin pulldown system were performed to confirm the interaction between miR-328 and TUG1. A gene expression array analysis using clinical samples and RT-qPCR suggested that enhancer of zeste homolog 2 (EZH2) was a target of miR-382 in PC. Results: In this study, we reported that TUG1 was overexpressed in PC tissues and cell lines, and high expression of TUG1 predicted poor prognosis. Further experiments revealed that overexpressed TUG1 promoted cell proliferation, migration and contributed to EMT formation, whereas silenced TUG1 led to opposing results. Additionally, luciferase reporter assays, an RIP assay and an RNA-pulldown assay demonstrated that TUG1 could competitively sponge miR-382 and thereby regulate EZH2. Conclusion: Collectively, these findings revealed that TUG1 functions as an oncogenic IncRNA that promotes tumor progression, at least partially, by functioning as an endogenous 'sponge' and competing for miR-382 binding to the miRNA target EZH2. (C) 2017 The Author(s) Published by S. Karger AG, Basel