Background

Long non-coding RNA (lncRNA) nuclear enriched abundant transcript 1 (NEAT1) has been documented to implicate in diverse tumor progression. However, the mechanism of NEAT1 in glioma was rarely reported.

Methods

The levels of NEAT1, microRNA-152-3p (miR-152-3p) and chaperonin containing TCP1 subunit 6A (CCT6A) in glioma tissues and cells were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The cell viability, apoptotic rate, the migrated and invaded abilities of A172 and U251 cells were evaluated via cell counting kit-8 (CCK-8), flow cytometry and Transwell assay, respectively. The mice xenograft model was constructed to further verify the effect of NEAT1. The interactions between miR-152-3p and NEAT1 or CCT6A were predicted by starBase v2.0 or TargetScan, then luciferase reporter assay, RNA immunoprecipitation (RIP) and RNA pull-down assay were performed to validate the interaction. The protein level of CCT6A was detected by Western blot assay.

Results

The levels of NEAT1, CCT6A were highly expressed, but miR-152-3p was decreased in glioma tissues and cells. NEAT1 depletion or miR-152-3p mimics suppressed cell viability, migrated and invaded abilities but induced apoptotic rate in A172 and U251 cells, while the introduction of CCT6A partly counteracted these impacts. In addition, NEAT1 silencing impeded xenograft tumor growth in vivo. MiR-152-3p was verified as a direct target of NEAT1 and directly targeted CCT6A. CCT6A expression was upregulated by NEAT1 and reversed by miR-152-3p.

Conclusion

NEAT1 enhanced glioma progression, partially through miR-152-3p/CCT6A pathway. The novel regulatory network might contribute to the diagnosis and treatment of glioma.

中文翻译：

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较长的非编码RNA NEAT1通过miR-152-3p / CCT6A途径促进人类神经胶质瘤肿瘤进展。

背景

长期的非编码RNA（lncRNA）核富集丰富的转录本1（NEAT1）已被证明与多种肿瘤进展有关。然而，关于神经胶质瘤中NEAT1的机制鲜有报道。

方法

通过定量实时聚合酶链反应（qRT-PCR）测量神经胶质瘤组织和细胞中NEAT1，microRNA-152-3p（miR-152-3p）和分子伴侣含有TCP1亚基6A（CCT6A）的水平。分别通过细胞计数试剂盒8（CCK-8），流式细胞仪和Transwell测定法评估A172和U251细胞的细胞活力，凋亡率，迁移能力和侵袭能力。构建小鼠异种移植模型以进一步验证NEAT1的作用。通过starBase v2.0或TargetScan预测miR-152-3p与NEAT1或CCT6A之间的相互作用，然后进行荧光素酶报告基因测定，RNA免疫沉淀（RIP）和RNA下拉测定来验证相互作用。通过蛋白质印迹法检测CCT6A的蛋白水平。

结果

NEAT1，CCT6A的水平高表达，但在胶质瘤组织和细胞中miR-152-3p降低。NEAT1耗竭或miR-152-3p模拟物抑制了细胞活力，迁移和侵袭能力，但诱导了A172和U251细胞的凋亡率，而CCT6A的引入部分抵消了这些影响。另外，NEAT1沉默抑制体内异种移植肿瘤的生长。验证了MiR-152-3p是NEAT1的直接靶标，也是CCT6A的直接靶标。CCT6A表达被NEAT1上调，而被miR-152-3p逆转。

结论

NEAT1增强了神经胶质瘤的进展，部分通过miR-152-3p / CCT6A途径。新的调节网络可能有助于神经胶质瘤的诊断和治疗。