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长链非编码 RNA DANCR 通过与鼻咽癌中的 NF90/NF45 复合物相互作用稳定 HIF-1α 并促进转移。

Long non-coding RNA DANCR stabilizes HIF-1α and promotes metastasis by interacting with NF90/NF45 complex in nasopharyngeal carcinoma.

机构信息

Sun Yat-sen University Cancer Center; State Key Laboratory of Oncology in South China; Collaborative Innovation Center of Cancer Medicine; Guangdong Key Laboratory of Nasopharyngeal Carcinoma Diagnosis and Therapy, No. 651 Dongfeng Road East, Guangzhou 510060, People's Republic of China.

出版信息

Theranostics. 2018 Nov 10;8(20):5676-5689. doi: 10.7150/thno.28538. eCollection 2018.

Abstract

Long noncoding RNAs (lncRNAs) play an important role in the development and progression of cancers. However, the clinical significances of lncRNAs and their functions and mechanisms in nasopharyngeal carcinoma (NPC) remain largely unclear. Quantitative RT-PCR was used to determine DANCR expression and Kaplan-Meier curves were used to evaluate its prognostic value. RNA sequencing followed by bioinformatic analysis was performed to determine the potential function of DANCR. and experiments were conducted to investigate its biological effects. DANCR-interacting proteins were identified by RNA pull-down assay followed by mass spectrometry and western blotting, and then confirmed by RNA immunoprecipitation (RIP) assays. Our previous microarray analysis identified a metastasis-associated lncRNA DANCR. Here, we found that DANCR was upregulated in NPC, especially in those with lymph lode metastasis, and its upregulation could predict poor survival. We then constructed a prognostic predictive model. RNA sequencing followed by bioinformatic analysis revealed that DANCR was responsible for NPC metastasis and hypoxia phenotype. Functional studies showed that DANCR promoted NPC cell invasion and metastasis and . Further investigation suggested that DANCR could increase HIF-1α mRNA stability through interacting with the NF90/NF45 complex. Additionally, overexpression of HIF-1α in DANCR knockdown cells restored its suppressive effects on NPC cell migration and invasion. Taken together, our results suggest that DANCR acts as a prognostic biomarker and increases HIF-1α mRNA stability by interacting with NF90/NF45, leading to metastasis and disease progression of NPC.

摘要

长链非编码 RNA(lncRNAs)在癌症的发生和发展中发挥着重要作用。然而,lncRNAs 的临床意义及其在鼻咽癌(NPC)中的功能和机制仍很大程度上不清楚。采用定量 RT-PCR 测定 DANCR 的表达,采用 Kaplan-Meier 曲线评估其预后价值。进行 RNA 测序及生物信息学分析,以确定 DANCR 的潜在功能。进行实验以研究其生物学效应。通过 RNA 下拉实验结合质谱和 Western blot 鉴定 DANCR 的相互作用蛋白,然后通过 RNA 免疫沉淀(RIP)实验进行验证。我们之前的微阵列分析确定了一个与转移相关的 lncRNA DANCR。在这里,我们发现 DANCR 在 NPC 中上调,特别是在那些有淋巴结转移的患者中,其上调可以预测不良预后。然后我们构建了一个预后预测模型。RNA 测序及生物信息学分析表明,DANCR 导致 NPC 转移和低氧表型。功能研究表明,DANCR 通过与 NF90/NF45 复合物相互作用促进 NPC 细胞侵袭和转移。进一步的研究表明,DANCR 通过与 NF90/NF45 复合物相互作用,增加 HIF-1α mRNA 的稳定性。此外,在 DANCR 敲低细胞中转染 HIF-1α 可以恢复其对 NPC 细胞迁移和侵袭的抑制作用。综上所述,我们的研究结果表明,DANCR 作为一个预后生物标志物,通过与 NF90/NF45 相互作用增加 HIF-1α mRNA 的稳定性,导致 NPC 的转移和疾病进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be06/6276287/ced9550c3051/thnov08p5676g001.jpg

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