miR-345-5p 的上调通过调节 ras 同源家族成员 A（RhoA）和 Rho/Rho 相关蛋白激酶（Rho/ROCK）通路抑制肺腺癌细胞的生长。

Upregulation of miR-345-5p suppresses cell growth of lung adenocarcinoma by regulating ras homolog family member A (RhoA) and Rho/Rho associated protein kinase (Rho/ROCK) pathway.

机构信息

Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230011, China.

Department of Respiratory Medicine, Anhui No. 2 Provincial People's Hospital, Clinical College of Anhui Medical University, Hefei, Anhui 230032, China.

出版信息

Chin Med J (Engl). 2021 Oct 14;134(21):2619-2628. doi: 10.1097/CM9.0000000000001804.

DOI:10.1097/CM9.0000000000001804

PMID:34748526

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8577671/

Abstract

BACKGROUND

Microribose nucleic acids (miRNAs) are implicated in the progression of lung adenocarcinoma. MicroRNA-345-5p (miR-345-5p) is a recently identified anti-oncogene in some human cancers, but its functional role and possible molecular mechanism in lung adenocarcinoma remain unknown. This study aimed to identify the biological function and underlying mechanism of miR-345-5p in lung adenocarcinoma cells.

METHODS

In this study, lung adenocarcinoma tissues and adjacent tissues were collected in the First Affiliated Hospital of Anhui Medical University between April 2016 and February 2017. The expression of miR-345-5p and ras homolog family member A (RhoA) in lung adenocarcinoma tissues and human lung adenocarcinoma cell lines (A549, H1650, PC-9, and H441) was detected by reverse transcription quantitative polymerase chain reaction analysis. Functional assays including colony formation, flow cytometry analysis, wound healing, and transwell assays were performed to assess the proliferation, apoptosis, migration, and invasion of lung adenocarcinoma cells. In addition, RNA pulldown and luciferase reporter assays were conducted to evaluate the relationship between miR-345-5p and RhoA. Difference between the two groups was analyzed with Student's t test, while that among multiple groups was analyzed with one-way analysis of variance.

RESULTS

MiR-345-5p expression displayed lower level in lung adenocarcinoma tissues (0.241 ± 0.095 vs.1.000 ± 0.233, t = 19.247, P < 0.001) and cell lines (F = 56.992, P < 0.001) than control tissues and cells. Functional experiments demonstrated that upregulation of miR-345-5p inhibited the malignant phenotypes of lung adenocarcinoma cells via suppressing cell proliferation, migration, invasion, and facilitating cell apoptosis. Additionally, RhoA was verified to be the downstream target of miR-345-5p. Expression of RhoA was downregulated by overexpression of miR-345-5p in PC-9 (0.321 ± 0.047 vs. 1.000 ± 0.127, t = 8.536, P < 0.001) and H1650 (0.398 ± 0.054 vs. 1.000 ± 0.156, t = 4.429, P = 0.011) cells. Rescue assays revealed that overexpression of RhoA rescued the suppressive effects of miR-345-5p upregulation on proliferation, migration, and invasion of lung adenocarcinoma cells. Further, miR-345-5p was found to regulate the Rho/Rho-associated protein kinase (ROCK) signaling pathway by downregulation of RhoA in lung adenocarcinoma cells.

CONCLUSIONS

MiR-345-5p plays a tumor suppressor role in lung adenocarcinoma cells by downregulating RhoA to inactivate the Rho/ROCK pathway.

摘要

背景

微小核糖核酸（miRNAs）参与肺腺癌的进展。miR-345-5p 是最近在一些人类癌症中被鉴定出的抑癌基因，但它在肺腺癌中的功能作用和可能的分子机制仍不清楚。本研究旨在鉴定 miR-345-5p 在肺腺癌细胞中的生物学功能和潜在机制。

方法

本研究于 2016 年 4 月至 2017 年 2 月期间在安徽医科大学第一附属医院收集肺腺癌组织和相邻组织。采用逆转录定量聚合酶链反应分析检测肺腺癌组织和人肺腺癌细胞系（A549、H1650、PC-9 和 H441）中 miR-345-5p 和 Ras 同源家族成员 A（RhoA）的表达。通过集落形成、流式细胞术分析、划痕愈合和 Transwell 分析等功能测定评估肺腺癌细胞的增殖、凋亡、迁移和侵袭。此外，进行 RNA 下拉和荧光素酶报告基因分析以评估 miR-345-5p 和 RhoA 之间的关系。采用 Student's t 检验分析两组间的差异，采用单因素方差分析分析多组间的差异。

结果

miR-345-5p 在肺腺癌组织（0.241 ± 0.095 比 1.000 ± 0.233，t = 19.247，P < 0.001）和细胞系（F = 56.992，P < 0.001）中的表达水平低于对照组织和细胞。功能实验表明，上调 miR-345-5p 通过抑制细胞增殖、迁移、侵袭和促进细胞凋亡来抑制肺腺癌细胞的恶性表型。此外，RhoA 被证实是 miR-345-5p 的下游靶标。在 PC-9（0.321 ± 0.047 比 1.000 ± 0.127，t = 8.536，P < 0.001）和 H1650（0.398 ± 0.054 比 1.000 ± 0.156，t = 4.429，P = 0.011）细胞中，过表达 miR-345-5p 下调 RhoA 的表达。挽救实验表明，过表达 RhoA 可挽救 miR-345-5p 上调对肺腺癌细胞增殖、迁移和侵袭的抑制作用。此外，在肺腺癌细胞中，miR-345-5p 通过下调 RhoA 来调节 Rho/Rho 相关蛋白激酶（ROCK）信号通路。