GATA2- miR-374a轴通过靶向circTADA2A/RORA轴促进血管平滑肌细胞增殖和迁移。

GATA2‑miR‑374a axis promotes vascular smooth muscle cells proliferation, migration via targeting circTADA2A/RORA axis.

作者信息

Tu Wenxian, Feng Meina, Zhou Qin, Wang Yunfeng, Wan Mingye, Gong Danqun, Li Jin, Du Yuanmin

机构信息

Department of Neurology, Wuhan Brain Hospital, General Hospital of the YANGTZE River Shipping, Wuhan, Hubei 430030, P.R. China.

出版信息

Exp Ther Med. 2024 Jul 8;28(3):357. doi: 10.3892/etm.2024.12646. eCollection 2024 Sep.

Abstract

Evidence has shown that microRNAs (miRNAs/miRs) play key roles in biological functions of vascular smooth muscle cells (VSMCs). However, the role of miR-374a in VSMCs remains to be elucidated. The present study aimed to explore the influence of miR-374a on VSMCs and its molecular mechanism. The expression level of miR-374a was measured by reverse transcription-quantitative (RT-q) PCR. MTT and Transwell assay were employed to assess the role of miR-374a in proliferation and migration of VSMCs. To order to determine miR-374a targets, a dual-luciferase reporter assay was conducted, which was further verified by rescue experiments. Chromatin Immunoprecipitation Assay and JASPAR databases were applied to explore the regulatory association between GATA binding protein 2 (GATA2) and miR-374a. Western blotting or RT-qPCR were employed to detect the protein expression levels of GATA2 or RAR-related orphan receptor A (RORA). The present study found that miR-374a was elevated in VSMCs following treatment with platelet-derived growth factor-BB (PDGF-BB) compared with that in control group. In addition, the results demonstrated that a higher expression of a miR-374a could promote proliferation and migration of VSMCs while miR-374a inhibitor suppressed the PDGF-BB-induced proliferation and migration of VSMCs . Furthermore, circTADA2A bound to miR-374a and then upregulated RORA expression, which resulted in inhibition in VSMCs proliferation and migration. On the other hand, the result indicated that GATA2 overexpression could augment the proliferation, migration of PDGF-bb-induced VSMCs, which could be rescued by miR-374a inhibitor. The findings suggested that the GATA2/circTADA2A-miR-374a axis promoted the proliferation and migration of VSMCs by targeting RORA, which were closely related to atherosclerosis (AS). Thus the results might offer a new therapeutic target for AS.

摘要

有证据表明,微小RNA(miRNA/miR)在血管平滑肌细胞(VSMC)的生物学功能中起关键作用。然而,miR-374a在VSMC中的作用仍有待阐明。本研究旨在探讨miR-374a对VSMC的影响及其分子机制。通过逆转录定量(RT-q)PCR检测miR-374a的表达水平。采用MTT和Transwell实验评估miR-374a在VSMC增殖和迁移中的作用。为了确定miR-374a的靶标,进行了双荧光素酶报告基因实验,并通过拯救实验进一步验证。应用染色质免疫沉淀实验和JASPAR数据库来探究GATA结合蛋白2(GATA2)与miR-374a之间的调控关系。采用蛋白质印迹法或RT-qPCR检测GATA2或视黄酸相关孤儿受体A(RORA)的蛋白表达水平。本研究发现,与对照组相比,血小板衍生生长因子-BB(PDGF-BB)处理后的VSMC中miR-374a升高。此外,结果表明,较高表达的miR-374a可促进VSMC的增殖和迁移,而miR-374a抑制剂可抑制PDGF-BB诱导的VSMC增殖和迁移。此外,环状TADA2A与miR-374a结合,进而上调RORA表达,导致VSMC增殖和迁移受到抑制。另一方面,结果表明,GATA2过表达可增强PDGF-bb诱导的VSMC的增殖和迁移,而miR-374a抑制剂可使其恢复。这些发现表明,GATA2/环状TADA2A-miR-374a轴通过靶向RORA促进VSMC的增殖和迁移,这与动脉粥样硬化(AS)密切相关。因此,这些结果可能为AS提供一个新的治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/26cc/11273358/914d903489ae/etm-28-03-12646-g00.jpg

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