Tan Xiangrong, Wang Peng, Lou Jianlin, Zhao Jiazheng
1Institute of Cancer and Basic Medicine (ICBM), Chinese Academy of Sciences, Hangzhou, Zhejiang China.
2Department of Head and Neck Surgery, Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou, Zhejiang China.
Cancer Cell Int. 2020 Apr 23;20:132. doi: 10.1186/s12935-020-01222-x. eCollection 2020.
Anaplastic thyroid carcinoma (ATC) is one of the most aggressive and lethal malignancies. Long non-coding RNAs (lncRNAs) are being found to play crucial roles in ATC progression. Herein, we focused on the role of nuclear paraspeckle assembly transcript 1 (NEAT1) on ATC progression under hypoxia and underlying mechanisms governing it.
The expression levels of NEAT1, miR-206 and miR-599 were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). Cell migration and invasion abilities were detected using transwell assays. Glucose consumption and lactate production were determined using a corresponding commercial assay kit. Western blot was performed to evaluate the level of hexokinase 2 (HK2). The targeted interplays between NEAT1 and miR-206 or miR-599 were confirmed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Xenograft model was established to observe the effect of NEAT1 on tumor growth in vivo.
Our data indicated that NEAT1 was highly expressed in ATC tissues and cells, and hypoxia induced NEAT1 expression in ATC cells. NEAT1 depletion repressed ATC cell migration, invasion and glycolysis under hypoxia. Mechanistically, NEAT1 acted as a molecular sponge of miR-206 and miR-599. Moreover, the repressive effects of NEAT1 knockdown on ATC cell migration, invasion and glycolysis under hypoxia were mediated by miR-206 or miR-599. Additionally, NEAT1 knockdown weakened tumor growth in vivo.
In conclusion, our study suggested that a low NEAT1 expression suppressed the migration, invasion, and glycolysis in ATC cells under hypoxia at least partially through modulating miR-206 and miR-599, providing new therapeutic strategies for ATC treatment.
间变性甲状腺癌（ATC）是最具侵袭性和致命性的恶性肿瘤之一。长链非编码RNA（lncRNAs）被发现在ATC进展中起关键作用。在此，我们聚焦于核旁斑组装转录本1（NEAT1）在缺氧条件下对ATC进展的作用及其潜在调控机制。
通过定量实时聚合酶链反应（qRT-PCR）评估NEAT1、miR-206和miR-599的表达水平。使用Transwell实验检测细胞迁移和侵袭能力。使用相应的商业检测试剂盒测定葡萄糖消耗和乳酸产生。进行蛋白质免疫印迹法以评估己糖激酶2（HK2）的水平。通过双荧光素酶报告基因和RNA免疫沉淀（RIP）实验证实NEAT1与miR-206或miR-599之间的靶向相互作用。建立异种移植模型以观察NEAT1对体内肿瘤生长的影响。
我们的数据表明，NEAT1在ATC组织和细胞中高表达，缺氧诱导ATC细胞中NEAT1表达。在缺氧条件下，NEAT1缺失抑制了ATC细胞的迁移、侵袭和糖酵解。机制上，NEAT1作为miR-206和miR-599的分子海绵。此外，缺氧条件下NEAT1敲低对ATC细胞迁移、侵袭和糖酵解的抑制作用是由miR-206或miR-599介导的。此外，NEAT1敲低减弱了体内肿瘤生长。
总之，我们的研究表明，低NEAT1表达至少部分通过调节miR-206和miR-599抑制缺氧条件下ATC细胞的迁移、侵袭和糖酵解，为ATC治疗提供了新的治疗策略。
