miR-647 通过调控 PTEN/PI3K/AKT 通路促进 ox-LDL 处理的血管平滑肌细胞的增殖和迁移。

MiR-647 promotes proliferation and migration of ox-LDL-treated vascular smooth muscle cells through regulating PTEN/PI3K/AKT pathway.

机构信息

Department of Cardiology, the Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian, China.

出版信息

Eur Rev Med Pharmacol Sci. 2019 Aug;23(16):7110-7119. doi: 10.26355/eurrev_201908_18756.

DOI:10.26355/eurrev_201908_18756

PMID:31486513

Abstract

OBJECTIVE

Aberrant microRNAs (miRNAs) play vital roles in various human diseases, including atherosclerosis (AS). MiR-647 expression was highly elevated in AS samples. Therefore, this study aimed at exploring the role and mechanism of miR-647 on AS progression.

PATIENTS AND METHODS

Human aorta vascular smooth muscle cells (HA-VSMCs) were treated with oxidized modified low-density lipoprotein (ox-LDL) to establish the AS model in vitro. The qRT-PCR assay was used to detect the expression of miR-647 and PTEN mRNA. The levels of PTEN protein, PI3K, AKT, p-PI3K, and p-AKT were measured using Western blot. Cell proliferation and migration were determined by Cell Counting Kit-8 (CCK-8) assay and transwell assay, respectively. The target of miR-647 was verified using the dual-luciferase reporter assay.

RESULTS

Our data supported that miR-647 was upregulated and PTEN was downregulated in the serum of AS patients and ox-LDL-treated HA-VSMCs. The proliferation and migration of ox-LDL-treated HA-VSMCs were promoted by miR-647 overexpression or PTEN knockdown, while they were suppressed following miR-647 depletion or high PTEN expression. Moreover, PTEN was a direct target of miR-647. PTEN antagonized miR-647-mediated regulatory effects on cell proliferation and migration. Additionally, the PI3K/AKT signaling pathway was involved in miR-647/PTEN-mediated regulation in ox-LDL-treated HA-VSMCs.

CONCLUSIONS

MiR-647 promoted the proliferation and migration of ox-LDL-treated HA-VSMCs at least partly by targeting the PTEN/PI3K/AKT pathway. Targeting miR-647 may be a promising method for AS treatment.

摘要

目的

异常表达的 microRNAs（miRNAs）在多种人类疾病中发挥着重要作用，包括动脉粥样硬化（AS）。miR-647 在 AS 样本中的表达水平显著升高。因此，本研究旨在探讨 miR-647 在 AS 进展中的作用和机制。

方法

采用氧化修饰的低密度脂蛋白（ox-LDL）处理人主动脉血管平滑肌细胞（HA-VSMCs）建立 AS 体外模型。采用 qRT-PCR 检测 miR-647 和 PTEN mRNA 的表达。采用 Western blot 检测 PTEN 蛋白、PI3K、AKT、p-PI3K 和 p-AKT 的水平。采用细胞计数试剂盒-8（CCK-8）检测细胞增殖，Transwell 检测细胞迁移。采用双荧光素酶报告基因检测验证 miR-647 的靶基因。

结果

我们的数据表明，AS 患者血清和 ox-LDL 处理的 HA-VSMCs 中 miR-647 表达上调，PTEN 表达下调。miR-647 过表达或 PTEN 敲低促进 ox-LDL 处理的 HA-VSMCs 的增殖和迁移，而 miR-647 耗竭或高表达 PTEN 则抑制其增殖和迁移。此外，PTEN 是 miR-647 的直接靶基因。PTEN 拮抗 miR-647 对细胞增殖和迁移的调节作用。此外，PI3K/AKT 信号通路参与了 ox-LDL 处理的 HA-VSMCs 中 miR-647/PTEN 介导的调节作用。