长链非编码 RNA LINC00511 通过海绵吸附 miR-126-5p/miR-218-5p 促进肺腺癌 COL1A1 介导的增殖和转移。

LncRNA LINC00511 promotes COL1A1-mediated proliferation and metastasis by sponging miR-126-5p/miR-218-5p in lung adenocarcinoma.

机构信息

Thoracic Surgery Department, Shengjing Hospital of China Medical University, Shenyang, 110136, Liaoning, China.

College of Basic Medical Science, China Medical University, No. 77 Puhe Road, Shenbei New District, Shenyang, 110001, Liaoning, China.

出版信息

BMC Pulm Med. 2022 Jul 16;22(1):272. doi: 10.1186/s12890-022-02070-3.

DOI:10.1186/s12890-022-02070-3

PMID:35842617

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9287882/

Abstract

BACKGROUND

Lung adenocarcinoma (LUAD) is currently the leading cause of cancer-related death worldwide. Long noncoding RNAs (lncRNAs) play key roles in tumor occurrence and development as crucial cancer regulators. The present study aimed to explore the molecular mechanism and regulatory network of Linc00511 in LUAD and to identify new potential therapeutic targets for LUAD.

METHODS

Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to determine the relative Linc00511 levels in LUAD tissues and cells. The proliferation, apoptosis, migration, and invasion abilities of LUAD cells were assessed by a Cell Counting Kit-8 (CCK-8) assay, a colony formation assay, flow cytometry, and a Transwell assay. Changes in hsa_miR-126-5p, hsa_miR-218-5p, and COL1A1 expression were analyzed using western blotting and RT-qPCR. Targeted binding between miR-126-5p/miR-218-5p and Linc00511 or COL1A1 was verified with a luciferase reporter system and confirmed by an RNA pulldown assay. The participation of the PI3K/AKT signaling pathway was confirmed via western blotting. Xenograft animal experiments were performed to detect the impact of Linc00511 on LUAD tumor growth in vivo.

RESULTS

In the present work, we observed that Linc00511 was upregulated in LUAD tissues and cells. Loss/gain-of-function experiments indicated that knockdown of Linc00511 significantly inhibited LUAD cell proliferation, migration and invasion and promoted LUAD cell apoptosis, whereas overexpression of Linc00511 showed the opposite effects. In addition, we determined that Linc00511 promoted COL1A1-mediated cell proliferation and cell motility by sponging miR-126-5p and miR-218-5p. Moreover, Linc00511 activated the PI3K/AKT signaling pathway through upregulation of COL1A1. Finally, silencing of Linc00511 inhibited LUAD tumor growth in vivo.

CONCLUSIONS

Linc00511 acts as a competing endogenous RNA to regulate COL1A1 by targeting miR-126-5p and miR-218-5p, thereby promoting the proliferation and invasion of LUAD cells.

摘要

背景

肺腺癌（LUAD）是目前全球癌症相关死亡的主要原因。长链非编码 RNA（lncRNA）作为重要的癌症调控因子，在肿瘤的发生和发展中发挥关键作用。本研究旨在探讨 Linc00511 在 LUAD 中的分子机制和调控网络，并为 LUAD 寻找新的潜在治疗靶点。

方法

采用实时定量聚合酶链反应（RT-qPCR）检测 LUAD 组织和细胞中 Linc00511 的相对水平。通过细胞计数试剂盒-8（CCK-8）检测、集落形成实验、流式细胞术和 Transwell 实验评估 LUAD 细胞的增殖、凋亡、迁移和侵袭能力。采用 Western blot 和 RT-qPCR 分析 hsa_miR-126-5p、hsa_miR-218-5p 和 COL1A1 表达的变化。通过荧光素酶报告系统和 RNA 下拉实验验证 miR-126-5p/miR-218-5p 与 Linc00511 或 COL1A1 之间的靶向结合，并通过 Western blot 验证 PI3K/AKT 信号通路的参与。进行异种移植动物实验检测 Linc00511 对 LUAD 肿瘤在体内生长的影响。

结果

在本研究中，我们观察到 Linc00511 在 LUAD 组织和细胞中上调。敲低 Linc00511 的功能实验表明，Linc00511 敲低显著抑制 LUAD 细胞的增殖、迁移和侵袭，并促进 LUAD 细胞凋亡，而过表达 Linc00511 则表现出相反的效果。此外，我们确定 Linc00511 通过海绵吸附 miR-126-5p 和 miR-218-5p 促进 COL1A1 介导的细胞增殖和细胞迁移。此外，Linc00511 通过上调 COL1A1 激活 PI3K/AKT 信号通路。最后，沉默 Linc00511 抑制 LUAD 肿瘤在体内的生长。