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长链非编码 RNA PVT1 通过作为分子海绵调节 miR-519d-3p 促进喉鳞状细胞癌的发展。

Long noncoding RNA PVT1 promotes laryngeal squamous cell carcinoma development by acting as a molecular sponge to regulate miR-519d-3p.

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, China.

Department of Otorhinolaryngology Head and Neck Surgery, Xi'an Jiaotong University, Xi'an, Shaanxi, China.

出版信息

J Cell Biochem. 2019 Mar;120(3):3911-3921. doi: 10.1002/jcb.27673. Epub 2018 Oct 10.

Abstract

OBJECTIVE

This study was designed to investigate the effects and mechanism of long noncoding RNA (lncRNA) PVT1 on cell migration, proliferation, and apoptosis of laryngeal squamous cell carcinoma (LSCC).

METHODS

We screened lncRNAs expression profiles in four pair LSCC and matched noncancerous tissues by microarray assay. The messenger RNA levels of PVT1 in tissues and cells were evaluated by quantitative real-time polymerase chain reaction analysis. StarBase website was used to predict the target miRNAs for PVT1. And the interaction between PVT1 and target miRNA-519d-3p in LSCC cells was analyzed using dual-luciferase reporter assay. MTT assay was used to investigate the cell viability. Cell counting assay was used to explore the cell proliferation. Annexin-V propidium iodide flow cytometry was used to examine the cell apoptosis, and transwell assay was used to investigate the effects of lncRNA PVT1 on cell migration.

RESULTS

PVT1 was significantly overexpressed in human LSCC tissues and several LSCC cell lines. Upregulation of lncRNA PVT1 markedly facilitated proliferation suppressed apoptosis and promoted cell migration in LSCC cells. We further demonstrated that silencing PVT1 strikingly suppressed proliferation, promoted apoptosis, and reduced migration in LSCC cells. Further bioinformatic analysis and dual-luciferase reporter assay revealed that PVT1 could function as an oncogenic transcript partly through sponging miR-519d-3p. Besides, mechanistic investigations indicated that PVT1 could promote cell and migration through interacting with miR-519d-3p.

CONCLUSION

LncRNA PVT1 is consistently overexpressed in human LSCC, and overexpression of lncRNA PVT1 contributes to the proliferation and migration of LSCC through inhibiting miR-519d-3p expression.

摘要

目的

本研究旨在探讨长链非编码 RNA(lncRNA)PVT1 对喉鳞状细胞癌(LSCC)细胞迁移、增殖和凋亡的影响及其机制。

方法

通过微阵列分析筛选出 4 对 LSCC 及配对非癌组织中的 lncRNA 表达谱。采用定量实时聚合酶链反应分析检测组织和细胞中 PVT1 的信使 RNA 水平。使用 StarBase 网站预测 PVT1 的靶 miRNAs。采用双荧光素酶报告基因分析检测 LSCC 细胞中 PVT1 与靶 miRNA-519d-3p 的相互作用。采用 MTT 法检测细胞活力。采用细胞计数法检测细胞增殖。采用 Annexin-V 碘化丙啶流式细胞术检测细胞凋亡,采用 Transwell 法检测 lncRNA PVT1 对细胞迁移的影响。

结果

PVT1 在人 LSCC 组织和几种 LSCC 细胞系中显著过表达。上调 lncRNA PVT1 显著促进 LSCC 细胞增殖,抑制凋亡,促进迁移。我们进一步证明,沉默 PVT1 可显著抑制 LSCC 细胞增殖,促进凋亡,减少迁移。进一步的生物信息学分析和双荧光素酶报告基因分析显示,PVT1 可部分通过海绵 miR-519d-3p 发挥致癌转录本的作用。此外,机制研究表明,PVT1 可通过与 miR-519d-3p 相互作用促进细胞和迁移。

结论

lncRNA PVT1 在人 LSCC 中持续过表达,过表达 lncRNA PVT1 通过抑制 miR-519d-3p 表达促进 LSCC 的增殖和迁移。

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