LINC00662 triggers malignant progression of chordoma by the activation of RNF144B via targeting miR-16-5p
LINC00662 triggers malignant progression of chordoma by the activation of RNF144B via targeting miR-16-5p
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DOI：
10.26355/eurrev_202002_20151
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发表时间：
2020-01-01
影响因子：
3.3
通讯作者：
Guo, X-L
中科院分区：
文献类型：
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作者：
Wang, C-B;Wang, Y.;Guo, X-L
目的：软骨瘤是一种罕见的恶性肿瘤，诊断和治疗困难，长链非编码RNA作为新的生物标志物在许多癌症中被频繁报道。本研究的目的是探讨长基因间非编码RNA 00662（LINC 00662）在脉络膜中的作用及其相关的作用机制。采用真实的时间-聚合酶链反应（qRT-PCR）检测microRNA-16- 5 p（miR-16- 5 p）。RNF 144 B、细胞增殖标志物（Cyclin D1和Ki 67）、上皮-间质转化（EMT）标志物（E-cadherin、Vimentin和N-cadherin）和糖酵解标志物[葡萄糖转运蛋白1（GLUT 1）、己糖激酶II（HK 2）]的蛋白水平。和乳酸脱氢酶A（LDHA）]。采用MTT法、集落形成实验和transwell实验分别观察细胞增殖、集落数、迁移和侵袭能力。通过糖酵解应激试验、葡萄糖消耗、乳酸产生和ATP产生来评价糖酵解进展。通过在线生物信息学工具starBase预测miR-16- 5 p与LINC 00662或RNF 144 B的关系，并通过双荧光素酶报告基因分析进行验证。结果：LINC 00662和RNF 144 B在脉络膜组织中表达异常上调，LINC 00662和RNF 144 B在脉络膜组织中表达异常上调。LINC 00662或RNF 144 B的敲低阻碍了脉络膜细胞的增殖、集落形成、侵袭、迁移、EMT和糖酵解。另外. RNF 144 B过表达逆转了LINC 00662敲低的作用。证实miR-16- 5 p是LINC 00662的靶点，并且miR-16 - 5 p可以靶向RNF 144 B。通过miR-16- 5 p建立LINC 00662与RNF 144 B之间的关系。另外。结论：LINC 00662作为miR-16- 5 p的海绵体，通过促进RNF 144 B参与脉络膜的恶性进展，提示LINC 00662是一个有希望的脉络膜治疗靶点。
OBJECTIVE: Chordoma is a rare malignant tumor difficult to diagnose and treat. Long non-coding RNAs acting as novel biomarkers are frequently reported in numerous cancers. The purpose of this study was to investigate the role of long intergenic non-coding RNA00662 (LINC00662) and its associated action mechanisms in chordoma.MATERIALS AND METHODS: The expression of LINC00662, Ring finger protein 144B (RNF144B). and microRNA-16-5p (miR-16-5p) was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The protein levels of RNF144B, cell proliferation markers (Cyclin D1 and Ki67), epithelial-mesenchymal transition (EMT) markers (E-cadherin, Vimentin and N-cadherin), and glycolysis markers [glucose transporter 1 (GLUT1), hexokinase II (HK2). and lactic dehydrogenase A (LDHA)] were determined by Western blot. Cell proliferation, the number of colonies, migration, and invasion were investigated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT), colony formation, and transwell assays, respectively. Glycolysis progress was evaluated by the glycolysis stress test, glucose consumption, lactate production, and ATP production. The relationship between miR-16-5p and LINC00662 or RNF144B was predicted by the online bioinformatics tool starBase and verified by Dual-Luciferase reporter assay. Xenograft tumor model was established to monitor the role of LINC00662 in vivo.RESULTS: LINC00662 and RNF144B were aberrantly upregulated in chordoma tissues. Knockdown of LINC00662 or RNF144B impeded proliferation, colony formation, invasion, migration, EMT, and glycolysis in chordoma cells. Besides. RNF144B overexpression reversed the role of LINC00662 knockdown. It was confirmed that miR-16-5p was a target of LINC00662, and miR16-5p could target RNF144B. The relationship between LINC00662 and RNF144B was established by miR-16-5p. In addition. LINC00662 stable knockdown inhibited tumor growth in vivo.CONCLUSIONS: LINC00662 participated in the malignant progression of chordoma by the promotion of RNF144B by acting as a sponge of miR-16-5p, suggesting that LINC00662 was a promising therapeutic target for chordoma.