长链非编码 RNA NEAT1 通过 miR-152-3p/CCT6A 通路促进人神经胶质瘤肿瘤进展。
Long non-coding RNA NEAT1 promotes human glioma tumor progression via miR-152-3p/CCT6A pathway.
机构信息
Department of Pathology, Shanghai Xuhui Central Hospital, Zhongshan-Xuhui Hospital, Fudan University, Shanghai Clincal Center, CAS, Shanghai, PR China.
Department of Neurosurgery, Shanghai Xuhui Central Hospital, Zhongshan-Xuhui Hospital, Fudan University, Shanghai Clincal Center, CAS, Shanghai, PR China.
出版信息
Neurosci Lett. 2020 Jul 27;732:135086. doi: 10.1016/j.neulet.2020.135086. Epub 2020 May 23.
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.
背景
长链非编码 RNA（lncRNA）核富集丰富转录物 1（NEAT1）已被证明参与多种肿瘤的进展。然而，NEAT1 在神经胶质瘤中的作用机制鲜有报道。
方法
采用实时定量聚合酶链反应（qRT-PCR）检测神经胶质瘤组织和细胞中 NEAT1、微小 RNA-152-3p（miR-152-3p）和热休克蛋白 10 家族成员 1A（HSP10A）的水平。通过细胞计数试剂盒-8（CCK-8）、流式细胞术和 Transwell 测定分别评估 A172 和 U251 细胞的细胞活力、凋亡率、迁移和侵袭能力。构建小鼠异种移植模型进一步验证 NEAT1 的作用。通过 starBase v2.0 或 TargetScan 预测 miR-152-3p 与 NEAT1 或 CCT6A 的相互作用，然后进行荧光素酶报告基因检测、RNA 免疫沉淀（RIP）和 RNA 下拉实验验证相互作用。Western blot 检测 CCT6A 蛋白水平。
结果
NEAT1、CCT6A 表达水平升高，miR-152-3p 表达降低，神经胶质瘤组织和细胞中。NEAT1 耗竭或 miR-152-3p 模拟物抑制 A172 和 U251 细胞的细胞活力、迁移和侵袭能力，但诱导细胞凋亡，而 CCT6A 的引入部分抵消了这些影响。此外，NEAT1 沉默抑制体内异种移植肿瘤生长。miR-152-3p 被证实是 NEAT1 的直接靶标，并直接靶向 CCT6A。NEAT1 上调 CCT6A 的表达，而 miR-152-3p 则逆转了 CCT6A 的表达。
结论
NEAT1 增强神经胶质瘤的进展，部分通过 miR-152-3p/CCT6A 通路。该新的调控网络可能有助于神经胶质瘤的诊断和治疗。
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