长链非编码 RNA FGD5-AS1 的沉默通过调节 miR-493-5p/DDX5 轴抑制非小细胞肺癌的进展。

Silencing of Long Non-Coding RNA FGD5-AS1 Inhibits the Progression of Non-Small Cell Lung Cancer by Regulating the miR-493-5p/DDX5 Axis.

机构信息

Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

Assisted Reproductive Center, Department of Obstetrics and Gynecology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

出版信息

Technol Cancer Res Treat. 2021 Jan-Dec;20:1533033821990007. doi: 10.1177/1533033821990007.

DOI:10.1177/1533033821990007

PMID:33550957

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7876571/

Abstract

BACKGROUND

Long non-coding RNA FGD5 antisense RNA 1 (FGD5-AS1), identified to be a carcinogenic lncRNA, exhibits a regulatory role in some malignancies including non-small cell lung cancer (NSCLC). The aim of the present research is to decipher the function and underlying mechanism of FGD5-AS1 in progression of NSCLC.

METHODS

Expression of FGD5-AS1, miR-493-5p and DEAD-box protein 5 (DDX5) in NSCLC tissues and cells was quantified utilizing qRT-PCR. Cell proliferation was assessed by CCK-8 method. Scratch healing test and Transwell assay were used for assaying cell migration and invasion. Expressions of DDX5 and epithelial-mesenchymal transition (EMT)-related proteins were examined by Western blot. Additionally, targeting relationships between FGD5-AS1 and miR-493-5p, miR-493-5p and DDX5 were verified by dual-luciferase reporter gene assay.

RESULTS

Expression of FGD5-AS1 in NSCLC tissues and cell lines was up-regulated. Expression of FGD5-AS1 was in association with enlarged tumor size and lymph node metastasis of the patients. Knockdown of FGD5-AS1 led to the inhibition of proliferation, migration, invasion and EMT of NSCLC cells. FGD5-AS1 directly targeted miR-493-5p, while DDX5 was the target of miR-493-5p in NSCLC cells. Additionally, FGD5-AS1 could positively regulate the expression of DDX5 via suppressing miR-493-5p.

CONCLUSION

FGD5-AS1 facilitates the proliferation, migration, invasion and EMT of NSCLC cells by sponging miR-493-5p and up-regulating DDX5.

摘要

背景

长链非编码 RNA FGD5 反义 RNA1（FGD5-AS1）被鉴定为致癌 lncRNA，在包括非小细胞肺癌（NSCLC）在内的一些恶性肿瘤中发挥调节作用。本研究旨在阐明 FGD5-AS1 在 NSCLC 进展中的功能和潜在机制。

方法

利用 qRT-PCR 定量检测 NSCLC 组织和细胞中 FGD5-AS1、miR-493-5p 和 DEAD 框蛋白 5（DDX5）的表达。通过 CCK-8 法评估细胞增殖。划痕愈合试验和 Transwell 测定用于检测细胞迁移和侵袭。通过 Western blot 检测 DDX5 和上皮-间充质转化（EMT）相关蛋白的表达。此外，通过双荧光素酶报告基因检测验证 FGD5-AS1 与 miR-493-5p、miR-493-5p 与 DDX5 之间的靶向关系。

结果

FGD5-AS1 在 NSCLC 组织和细胞系中的表达上调。FGD5-AS1 的表达与患者肿瘤体积增大和淋巴结转移有关。FGD5-AS1 敲低导致 NSCLC 细胞增殖、迁移、侵袭和 EMT 受到抑制。FGD5-AS1 可直接靶向 miR-493-5p，而 DDX5 是 NSCLC 细胞中 miR-493-5p 的靶基因。此外，FGD5-AS1 可通过抑制 miR-493-5p 正向调节 DDX5 的表达。