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长链非编码 RNA WTAPP1 通过调控 microRNA-592 促进喉癌细胞的增殖。

LncRNA WTAPP1 promotes proliferation of laryngeal carcinoma cells through regulating microRNA-592.

机构信息

Department of Clinical Laboratory, The Second Children & Women's Healthcare of Jinan City, Jinan, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9532-9540. doi: 10.26355/eurrev_202009_23038.

Abstract

OBJECTIVE

The aim of this work was to investigate the mechanism by which long non-coding RNA (lncRNA) WTAPP1 promotes the malignant progression of laryngeal cancer.

PATIENTS AND METHODS

In this study, quantitative real-time polymerase chain reaction (qRT-PCR) examined the expression of lncRNA WTAPP1 in 49 pairs of tumor tissue specimens and paracancerous normal ones collected from laryngeal cancer patients. Subsequently, in the laryngeal squamous cell carcinoma cell lines AMC-HN-8 and Hep-2, WTAPP1 overexpression and knockdown vectors were constructed using lentivirus, and cell counting kit-8 (CCK-8), cell colony formation and 5-ethynyl-2'-deoxyuridine (EdU) assays were carried out to analyze the impact of lncRNA WTAPP1 on the function of laryngeal cancer cells. Finally, Luciferase reporting assay and recovery experiments were carried out to further explore whether lncRNA WTAPP1 has an impact on the malignant progression of laryngeal cancer via modulating microRNA-592.

RESULTS

QRT-PCR results revealed a significantly higher expression of lncRNA WTAPP1 in tumor tissues of patients with laryngeal cancer than that in adjacent normal ones. Compared with patients with low expression of WTAPP1, those with higher expression had a more advanced pathological stage. Meanwhile, the proliferation ability of cells in sh-WTAPP1 group was remarkably attenuated when compared with that in sh-NC group. In addition, microRNA-592 and WTAPP1 mRNA levels were found negatively correlated in laryngeal carcinoma tissue specimens. Luciferase reporter gene assay indicated that WTAPP1 can be targeted by microRNA-592 through certain binding sites. Moreover, we demonstrated through some recovery experiments that WTAPP1 can indeed serve as an oncogene accelerating the malignant progression of laryngeal cancer through the modulation of microRNA-592.

CONCLUSIONS

LncRNA WTAPP was markedly highly expressed both in laryngeal carcinoma tissues and cell lines, which was also found to be closely relevant to the pathological stage of laryngeal cancer patients. Additionally, lncRNA WTAPP1 is able to enhance the proliferation capacity of laryngeal carcinoma cells via regulating microRNA-592.

摘要

目的

本研究旨在探讨长链非编码 RNA(lncRNA)WTAPP1 促进喉癌恶性进展的机制。

患者与方法

采用实时定量聚合酶链反应(qRT-PCR)检测 49 对喉癌患者肿瘤组织和癌旁正常组织中 lncRNA WTAPP1 的表达。随后,在喉鳞状细胞癌细胞系 AMC-HN-8 和 Hep-2 中,利用慢病毒构建 WTAPP1 过表达和敲低载体,通过细胞计数试剂盒-8(CCK-8)、细胞集落形成和 5-乙炔基-2'-脱氧尿苷(EdU)实验分析 lncRNA WTAPP1 对喉癌细胞功能的影响。最后,通过荧光素酶报告基因检测和恢复实验进一步探讨 lncRNA WTAPP1 是否通过调节 microRNA-592 对喉癌的恶性进展产生影响。

结果

qRT-PCR 结果显示,喉癌患者肿瘤组织中 lncRNA WTAPP1 的表达明显高于癌旁正常组织。与 WTAPP1 低表达的患者相比,高表达的患者具有更晚期的病理分期。同时,sh-WTAPP1 组细胞的增殖能力明显低于 sh-NC 组。此外,在喉癌组织标本中发现 microRNA-592 和 WTAPP1 mRNA 水平呈负相关。荧光素酶报告基因实验表明,WTAPP1 可以通过特定的结合位点被 microRNA-592 靶向。此外,通过一些恢复实验表明,WTAPP1 可以通过调节 microRNA-592 作为癌基因加速喉癌的恶性进展。

结论

lncRNA WTAPP 在喉癌组织和细胞系中均明显高表达,且与喉癌患者的病理分期密切相关。此外,lncRNA WTAPP1 通过调节 microRNA-592 增强喉癌细胞的增殖能力。

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