环状 RNA EIF4G2 通过靶向 miR-26a 对急性心肌梗死的调控机制。
Regulatory Mechanism of circEIF4G2 Targeting miR-26a in Acute Myocardial Infarction.
机构信息
Department of Cardiology, Panyu Central Hospital, Guangzhou 511400, China.
Cardiovascular Institute of Panyu District, Guangzhou 511400, China.
出版信息
J Healthc Eng. 2022 Mar 16;2022:5308372. doi: 10.1155/2022/5308372. eCollection 2022.
BACKGROUND
Acute myocardial infarction (AMI) involves a series of complex cellular and molecular events, including circular RNAs (circRNAs), microRNAs (miRNAs) and other noncoding RNAs.
OBJECTIVE
In this study, the regulation mechanism of circEIF4G2 acting on miR-26a on HUVECs (Human Umbilical Vein Endothelial Cells) proliferation, cell cycle and angiogenesis ability was mainly explored in the vascular endothelial growth factor induced (VEGF-induced) angiogenesis model.
METHODS
VEGF induced HUVECs angiogenesis model was constructed, and the expression of circEIF4G2 and miR-26a in VEGF model was detected by qRT-PCR. When circEIF4G2 and miR-26a were knocked down or overexpressed in HUVECs, qRT-PCR was used to detect the expression of circEIF4G2 and miR-26a, CCK-8 was used to detect cell proliferation, flow cytometry was used to detect the cell cycle transition of HUVECs, and cell formation experiment was used to detect the ability of angiogenesis. MiRanda database and Targetscan predicted the binding site of circEIF4G2 and miR-26a, lucifase reporting assay and RNA pull down assay verified the interaction between circEIF4G2 and miR-26a.
RESULTS
After HUVECs were treated with VEGF, circEIF4G2 was significantly upregulated. After circEIF4G2 was knocked down, the proliferation and angiogenesis of HUVECs cells were decreased, and the process of cell cycle G0/G1 phase was blocked. The overexpression of miR-26a reduced the proliferation and angiogenesis of HUVECs cells and blocked the cell cycle progression of G0/G1 phase. Double lucifase reporter gene assay verified that circEIF4G2 could directly interact with miR-26a through the binding site, and RNA Pull down assay further verified the interaction between circEIF4G2 and miR-26a. When circEIF4G2 and miR-26a were knocked down simultaneously in HUVECs, it was found that knocking down miR-26a could reverse the inhibition of circEIF4G2 on cell proliferation, cycle and angiogenesis.
CONCLUSION
In the VEGF model, circEIF4G2 was highly expressed and miR-26a was low expressed. MiR-26a regulates HUVECs proliferation, cycle and angiogenesis by targeting circEIF4G2.
背景
急性心肌梗死（AMI）涉及一系列复杂的细胞和分子事件，包括环状 RNA（circRNA）、微小 RNA（miRNA）和其他非编码 RNA。
目的
本研究主要探讨血管内皮生长因子诱导（VEGF 诱导）血管生成模型中 circEIF4G2 对 miR-26a 作用于 HUVECs（人脐静脉内皮细胞）增殖、细胞周期和血管生成能力的调控机制。
方法
构建 VEGF 诱导的 HUVECs 血管生成模型，通过 qRT-PCR 检测 VEGF 模型中 circEIF4G2 和 miR-26a 的表达。当 HUVECs 中敲低或过表达 circEIF4G2 和 miR-26a 时，通过 qRT-PCR 检测 circEIF4G2 和 miR-26a 的表达，通过 CCK-8 检测细胞增殖，通过流式细胞术检测 HUVECs 细胞周期的转变，通过细胞形成实验检测血管生成能力。通过 MiRanda 数据库和 Targetscan 预测 circEIF4G2 和 miR-26a 的结合位点，通过荧光素酶报告基因测定和 RNA 下拉实验验证 circEIF4G2 和 miR-26a 之间的相互作用。
结果
VEGF 处理 HUVECs 后，circEIF4G2 明显上调。敲低 circEIF4G2 后，HUVECs 细胞的增殖和血管生成能力降低，细胞周期 G0/G1 期阻滞。过表达 miR-26a 可降低 HUVECs 细胞的增殖和血管生成能力，并阻滞 G0/G1 期细胞周期进程。双荧光素酶报告基因测定验证 circEIF4G2 可通过结合位点直接与 miR-26a 相互作用，RNA 下拉实验进一步验证了 circEIF4G2 与 miR-26a 之间的相互作用。当 HUVECs 中同时敲低 circEIF4G2 和 miR-26a 时，发现敲低 miR-26a 可逆转 circEIF4G2 对细胞增殖、周期和血管生成的抑制作用。
结论
在 VEGF 模型中，circEIF4G2 高表达，miR-26a 低表达。miR-26a 通过靶向 circEIF4G2 调节 HUVECs 的增殖、周期和血管生成。
Zotero 插件