长链非编码RNA CRNDE通过miR-545-5p/CCND2轴加剧鼻咽癌进展。

Long non-coding RNA CRNDE exacerbates NPC advancement mediated by the miR-545-5p/CCND2 axis.

作者信息

Ge Sichen, Jiang Chengyi, Li Min, Cheng Zhongqiang, Feng Xiaojia

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, The First Affiliated Hospital of Bengbu Medical College, Bengbu, 233000, Anhui, China.

Department of Surgical Oncology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, 233000, Anhui, China.

出版信息

Cancer Cell Int. 2021 Dec 4;21(1):650. doi: 10.1186/s12935-021-02348-2.

DOI:10.1186/s12935-021-02348-2

PMID:34863152

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8645150/

Abstract

BACKGROUND

Previous studies indicated CRNDE to have a pivotal part within tumorigenesis. Notwithstanding, precise details on CRNDE activities within NPC are still uncertain. The investigation described in this article served to focus in greater depth on the mechanistics regarding CRNDE, together with all associated regulatory networks, on nasopharyngeal carcinoma (NPC) and its treatment possibilities.

METHODS

Quantitative real-time polymerase chain reaction (RT-qPCR) analyzed CRNDE, miR-545-5p and CCND2 expression within NPCs and representative cell lineages. CCK-8 cell counting-, EdU-, wound-healing-/transwell-assays analyzed cellular proliferation, migrative, together with invasive properties. Apoptosis/cell cycle progression were scrutinized through flow cytometry. Dual-luciferase reporter assays validated CRNDE/miR-545-5p/CCND2 interplay. Proteomic expression of apoptosis-related protein, EMT-related protein and CCND2 protein were evaluated through Western blotting. In addition, Ki67 expression was evaluated through immunohistochemical staining. The effect of CRNDE in vivo was assessed by nude murine xenograft model studies.

RESULTS

This study demonstrated up-regulated expression of CRNDE and CCND2 within NPC tissues/cell lines. Meanwhile, miR-545-5p was down-regulated. CRNDE knock-down or miR-545-5p over-expression drastically reduced NPC proliferative, migrative and invasive properties, promoted apoptosis/altered cell cycle, and inhibited CCND2 expression. However, miR-545-5p down-regulation had opposing effects. All inhibiting functions generated by CRNDE down-regulation upon NPC progression could be counterbalanced or synergistically exacerbated, depending on miR-545-5p down-regulation or up-regulation, respectively. Multiple-level investigations revealed CRNDE to serve as a sponge for miR-545-5p, and can target CCND2 within NPCs.

CONCLUSIONS

CRNDE increases CCND2 expression by competitive binding with miR-545-5p, thus accelerating the development of NPC. This provides potential therapeutic targets and prognostic markers against NPC.

摘要

背景

先前的研究表明CRNDE在肿瘤发生过程中起关键作用。尽管如此，CRNDE在鼻咽癌（NPC）中的具体活动细节仍不明确。本文所述的研究旨在更深入地关注CRNDE的作用机制及其所有相关调控网络，以及鼻咽癌及其治疗可能性。

方法

采用定量实时聚合酶链反应（RT-qPCR）分析鼻咽癌组织和代表性细胞系中CRNDE、miR-545-5p和CCND2的表达。通过CCK-8细胞计数、EdU、伤口愈合和Transwell实验分析细胞增殖、迁移和侵袭特性。通过流式细胞术检测细胞凋亡和细胞周期进程。双荧光素酶报告基因实验验证CRNDE/miR-545-5p/CCND2之间的相互作用。通过蛋白质印迹法评估凋亡相关蛋白、上皮-间质转化（EMT）相关蛋白和CCND2蛋白的蛋白质组表达。此外，通过免疫组织化学染色评估Ki67的表达。通过裸鼠异种移植模型研究评估CRNDE在体内的作用。

结果

本研究表明，CRNDE和CCND2在鼻咽癌组织和细胞系中表达上调。同时，miR-545-5p表达下调。CRNDE敲低或miR-545-5p过表达显著降低鼻咽癌的增殖、迁移和侵袭特性，促进细胞凋亡/改变细胞周期，并抑制CCND2表达。然而，miR-545-5p下调则产生相反的效果。CRNDE下调对鼻咽癌进展产生的所有抑制作用，分别取决于miR-545-5p的下调或上调，可能会被抵消或协同增强。多层次研究表明，CRNDE可作为miR-545-5p的海绵，并可在鼻咽癌中靶向CCND2。