环状 RNA ACAP2 通过海绵吸附 miR-29 诱导心肌梗死后心肌细胞凋亡。
CircRNA ACAP2 induces myocardial apoptosis after myocardial infarction by sponging miR-29.
机构信息
Department of Internal and Cardiovascular Medicine, Yantaishan Hospital, Yantai, China.
Department of Internal and Cardiovascular Medicine, Jiangsu Taizhou People's Hospital, Taizhou, China.
出版信息
Minerva Med. 2022 Feb;113(1):128-134. doi: 10.23736/S0026-4806.20.06600-8. Epub 2020 May 13.
BACKGROUND
The aim of this study was to investigate the effect of circular RNA circ ACAP2 on apoptotic phenotype of rat cardiomyocytes and its molecular mechanism.
METHODS
Left anterior descending ligation was used to establish a rat myocardial infarction (MI) model, and real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of circ ACAP2 in apoptotic rat cardiomyocytes. The stability of circ ACAP2 was tested by actinomycin D and RNase R. H9C2 cells were infected by recombinant circ ACAP2 adenovirus (rAd-circ ACAP2), and the expression of apoptotic related genes Bax and BCL-2 in H9C2 was detected. Double luciferase reporter gene experiment, RNA antisense purification experiment and RNA Pull-Down experiments verified the binding effect of circ ACAP2 and miR-29.
RESULTS
RT-qPCR results showed that the expression of circ ACAP2 was increased in cardiomyocytes of MI rats. Actinomycin D and RNase R experiment confirmed that circ ACAP2 had better stability and resistance to RNase R degradation than its host gene ACAP2 mRNA. Overexpression of circ ACAP2 promotes apoptosis. The double luciferase reporter gene assay, RNA antisense purification assay and RNA Pulldown assay consistently confirmed the specific binding effect between circ ACAP2 and miR-29, and circ ACAP2 promoted the apoptotic phenotype in rats.
CONCLUSIONS
The expression of circ ACAP2 increased in cardiomyocytes after MI and promoted apoptosis by binding to miR-29.
背景
本研究旨在探讨环状 RNA circ ACAP2 对大鼠心肌细胞凋亡表型的影响及其分子机制。
方法
采用左冠状动脉前降支结扎法建立大鼠心肌梗死（MI）模型，实时定量聚合酶链反应（RT-qPCR）检测凋亡大鼠心肌细胞中 circ ACAP2 的表达。用放线菌素 D 和 RNase R 检测 circ ACAP2 的稳定性。用重组 circ ACAP2 腺病毒（rAd-circ ACAP2）感染 H9C2 细胞，检测 H9C2 中凋亡相关基因 Bax 和 BCL-2 的表达。双荧光素酶报告基因实验、RNA 反义纯化实验和 RNA 下拉实验验证了 circ ACAP2 与 miR-29 的结合效应。
结果
RT-qPCR 结果显示，MI 大鼠心肌细胞中 circ ACAP2 的表达增加。放线菌素 D 和 RNase R 实验证实，circ ACAP2 比其宿主基因 ACAP2mRNA 具有更好的稳定性和对 RNase R 降解的抗性。circ ACAP2 的过表达促进细胞凋亡。双荧光素酶报告基因实验、RNA 反义纯化实验和 RNA 下拉实验一致证实了 circ ACAP2 与 miR-29 之间的特异性结合效应，circ ACAP2 通过与 miR-29 结合促进大鼠的凋亡表型。
结论
MI 后心肌细胞中 circ ACAP2 的表达增加，并通过与 miR-29 结合促进细胞凋亡。
Zotero 插件