环状 RNA hsa_circ_0000317 通过调控 microRNA-494-3p/染色体 10 上缺失的磷酸酶及张力蛋白同源物轴抑制非小细胞肺癌进展。

Circular RNA hsa_circ_0000317 inhibits non-small cell lung cancer progression through regulating microRNA-494-3p/phosphatase and tensin homolog deleted on chromosome 10 axis.

机构信息

Department of Cardiothoracic Surgery, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Hubei, China.

出版信息

Clinics (Sao Paulo). 2022 Jul 30;77:100086. doi: 10.1016/j.clinsp.2022.100086. eCollection 2022.

DOI:10.1016/j.clinsp.2022.100086

PMID:35917658

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9344349/

Abstract

BACKGROUND

Circular RNA (circRNA), a group of non-coding RNA, is pivotal in the progression of various cancers, including Non-Small Cell Lung Cancer (NSCLC). Some circRNAs have been reported to be implicated in the progression of NSCLC, however, the function and molecular mechanism of hsa_circ_0000317 (circ_0000317) in NSCLC have not been fully understood.

METHODS

The significantly differentially expressed circRNA in NSCLC tissues, circ_0000317, was screened out by microarray. Circ_0000317, microRNA(miR)-494-3p and Phosphatase and Tensin Homolog Deleted on Chromosome 10 (PTEN) expressions in NSCLC tissues were respectively probed by quantitative real-time polymerase chain reaction and western blot assay. MTT and Transwell assays were adopted to examine the growth, migration, and invasion of NSCLC cells. Bioinformatics, luciferase reporter gene assay, RNA immunoprecipitation, and RNA pull-down assay were conducted to probe the relationships among circ_0000317, miR-494-3p, and PTEN.

RESULTS

Circ_0000317 expression level was reduced in NSCLC tissues and cell lines. Circ_0000317 expression in NSCLC patients was associated with TNM stage and lymphatic metastasis. Circ_0000317 overexpression restrained the proliferation, migration, and invasion of NSCLC cells, but co-transfection of miR-494-3p mimics partially reversed this effect. In addition, circ_0000317, was identified as a competitive endogenous RNA, which could sponge miR-494-3p to increase PTEN expression and activate PI3K/AKT pathway.

CONCLUSION

Circ_0000317, inhibits NSCLC progression via modulating miR-494-3p/PTEN/PI3K/AKT pathway.

摘要

背景

环状 RNA（circRNA）是一组非编码 RNA，在多种癌症的进展中起着关键作用，包括非小细胞肺癌（NSCLC）。已有研究报道，一些 circRNA 参与了 NSCLC 的进展，但 hsa_circ_0000317（circ_0000317）在 NSCLC 中的功能和分子机制尚未完全阐明。

方法

通过微阵列筛选出 NSCLC 组织中差异表达显著的 circRNA，circ_0000317。采用实时定量聚合酶链反应和 Western blot 检测 NSCLC 组织中 circ_0000317、miR-494-3p 和 10 号染色体缺失的磷酸酶和张力蛋白同源物（PTEN）的表达。采用 MTT 和 Transwell 实验检测 NSCLC 细胞的生长、迁移和侵袭。通过生物信息学、荧光素酶报告基因检测、RNA 免疫沉淀和 RNA 下拉实验探讨 circ_0000317、miR-494-3p 和 PTEN 之间的关系。

结果

circ_0000317 在 NSCLC 组织和细胞系中的表达水平降低。NSCLC 患者的 circ_0000317 表达与 TNM 分期和淋巴转移相关。circ_0000317 过表达抑制 NSCLC 细胞的增殖、迁移和侵袭，但共转染 miR-494-3p 模拟物部分逆转了这一效应。此外，circ_0000317 被鉴定为一种竞争性内源性 RNA，可通过海绵吸附 miR-494-3p 增加 PTEN 表达并激活 PI3K/AKT 通路。