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长链非编码 RNA DANCR 通过调节 miR-1225-3p/ErbB2 信号促进非小细胞肺癌细胞转移。

Long non-coding RNA DANCR promoted non-small cell lung cancer cells metastasis via modulating of miR-1225-3p/ErbB2 signal.

机构信息

Department of Thoracic Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, China.

出版信息

Eur Rev Med Pharmacol Sci. 2021 Jan;25(2):758-769. doi: 10.26355/eurrev_202101_24637.

Abstract

OBJECTIVE

Currently, we aimed to illustrate the role of lncRNA differentiation antagonizing non-protein coding RNA (DANCR) and erb-b2 receptor tyrosine kinase 2 (ErbB2) in non-small cell lung cancer (NSCLC).

PATIENTS AND METHODS

Expression of DANCR, microRNA-1225-3p (miR-1225-3p) and ErbB2 mRNA was evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) assays. The clinical value of DANCR was checked by a ROC curve analysis, a Kaplan-Meier analysis and a Pearson Chi-Square test. Transwell chamber assays were performed to determine the migration and invasion ability changes of SPCA1 and A549 cells. The protein expression of ErbB2 was tested by Western blot assays. The targeted binding effect between miR-1225-3p and DANCR or ErbB2 was confirmed by a Dual-Luciferase reporter assay and an RNA pull-down assay, respectively.

RESULTS

In the current study, it was found that DANCR was upregulated and correlated with poor prognosis in patients with NSCLC. DANCR promoted NSCLC cells migration and invasion via upregulation of ErbB2. DANCR regulated ErbB2 at posttranscriptional level. Mechanically, it was illustrated that miR-1225-3p negatively regulated ErbbB2 and it-mediated migration and invasion via directly targeting in NSCLC cells. Meanwhile, it was showed that DANCR interacted with miR-1225-3p in a reciprocal suppression manner. Even further, through a RIP assay and a luciferase assay, we showed that DANCR interacted with miR-1225-3p through a microRNA response element (MRE-1225-3p) via directly binding. Finally, it was demonstrated that DANCR served as a miR-1225-3p sponge to promote ErbB2 expression and to facilitate ErbB2-mediated migration and invasion in NSCLC cells.

CONCLUSIONS

In the current study, it was illustrated that DNACR promoted ErbB2-mediated migration and invasion via working as a ceRNA of miR-1225-3p in NSCLC cells.

摘要

目的

目前,我们旨在阐明长链非编码 RNA 分化拮抗非蛋白编码 RNA(DANCR)和表皮生长因子受体 2(ErbB2)在非小细胞肺癌(NSCLC)中的作用。

患者和方法

通过实时定量聚合酶链反应(qRT-PCR)检测 DANCR、microRNA-1225-3p(miR-1225-3p)和 ErbB2 mRNA 的表达。通过 ROC 曲线分析、Kaplan-Meier 分析和 Pearson Chi-Square 检验检查 DANCR 的临床价值。通过 Transwell 室测定法测定 SPCA1 和 A549 细胞迁移和侵袭能力的变化。通过 Western blot 测定法检测 ErbB2 的蛋白表达。通过双荧光素酶报告基因测定法和 RNA 下拉测定法分别证实 miR-1225-3p 与 DANCR 或 ErbB2 之间的靶向结合效应。

结果

在本研究中,发现 DANCR 在 NSCLC 患者中上调,并与不良预后相关。DANCR 通过上调 ErbB2 促进 NSCLC 细胞迁移和侵袭。DANCR 在转录后水平调节 ErbB2。从机制上讲,在 NSCLC 细胞中,miR-1225-3p 负调控 ErbbB2,并通过直接靶向调节其迁移和侵袭。同时,表明 DANCR 以相互抑制的方式与 miR-1225-3p 相互作用。甚至更进一步,通过 RIP 测定法和荧光素酶测定法,我们表明 DANCR 通过直接结合通过 microRNA 反应元件(MRE-1225-3p)与 miR-1225-3p 相互作用。最后,证明 DANCR 作为 miR-1225-3p 的海绵体通过促进 ErbB2 表达并促进 NSCLC 细胞中 ErbB2 介导的迁移和侵袭来发挥作用。

结论

在本研究中,阐明了 DANCR 通过作为 miR-1225-3p 的 ceRNA 在 NSCLC 细胞中促进 ErbB2 介导的迁移和侵袭。

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