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Circ_ROBO2/miR-186-5p/TRIM14 axis regulates oxidized low-density lipoprotein-induced cardiac microvascular
Qinghu Ye1, Changlin Ju2, Zhou Ye1
1Department of Cardiology, The First People's Hospital of Hangzhou Lin'an District, Ward 1, Hangzhou 311300, China.
Insights
Circular RNA ROBO2 (circ_ROBO2) is upregulated in coronary artery disease (CAD) and promotes cell injury by interacting with microRNA-186-5p and regulating TRIM14. This suggests circ_ROBO2 as a potential therapeutic target for CAD.
Area of Science:
- Cardiovascular Biology
- Molecular Biology
- Genetics
Background:
- Coronary artery disease (CAD) poses a significant mortality risk, primarily due to atherosclerosis.
- Circular RNAs (circRNAs) are increasingly recognized for their regulatory roles in cardiovascular diseases.
- The specific function of circ_ROBO2 in the context of CAD requires further elucidation.
Purpose of the Study:
- To investigate the role and mechanism of circ_ROBO2 in the pathogenesis of coronary artery disease (CAD).
- To explore the interaction between circ_ROBO2, microRNA-186-5p, and TRIM14 in cardiac microvascular endothelial cells (CMECs) under oxidative stress.
Main Methods:
- Established a cellular model of CAD using oxidized low-density lipoprotein (ox-LDL)-stimulated CMECs.
- Quantified RNA and protein expression using RT-qPCR and Western blot.
- Assessed cell proliferation, apoptosis, and angiogenesis using EdU, CCK-8, flow cytometry, and Matrigel assays.
- Verified molecular interactions via dual-luciferase reporter and RNA-pull down assays.
Main Results:
- Circ_ROBO2 expression was elevated in CAD patients and ox-LDL-treated CMECs.
- Ox-LDL induced CMEC injury, suppressed proliferation and angiogenesis, and promoted apoptosis, partly via circ_ROBO2 upregulation.
- Circ_ROBO2 acted as a molecular sponge for microRNA-186-5p (miR-186-5p), and knockdown of circ_ROBO2 protected CMECs from ox-LDL-induced damage.
- TRIM14 was identified as a target of miR-186-5p, and circ_ROBO2 positively regulated TRIM14 by sponging miR-186-5p.
Conclusions:
- Circ_ROBO2 exacerbates ox-LDL-induced injury in CMECs by sponging miR-186-5p and consequently regulating TRIM14.
- The circ_ROBO2/miR-186-5p/TRIM14 axis presents a potential therapeutic strategy for treating CAD.
Background:
Coronary artery disease (CAD) is one of the main risks of death, which is mainly caused by coronary arteries arteriosclerosis. Circular RNAs (circRNAs) have shown important regulatory roles in cardiovascular diseases. We amid to explore the role of circ_ROBO2 in CAD.
Methods:
Cardiac microvascular endothelial cells (CMECs) stimulated by oxidized low-density lipoprotein (ox-LDL) were served as the cellular model of CAD. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot assay were performed to detect RNA levels and protein levels, respectively. Cell proliferation was assessed by 5-ethynyl-2'-deoxyuridine (EdU) assay and Cell Counting Kit-8 (CCK-8) assay. Flow cytometry was employed for measuring cell apoptosis. Matrigel tube formation assay was used to evaluate angiogenesis ability. The intermolecular interaction was predicted by bioinformatics analysis and verified by dual-luciferase reporter and RNA-pull down assays.
Results:
The expression of circ_ROBO2 was upregulated in CAD patients and ox-LDL-induced CMECs. Treatment of ox-LDL suppressed cell proliferation and angiogenic ability as well as promoted the apoptosis of CMECs partly by upregulating circ_ROBO2. MicroRNA-186-5p (miR-186-5p) was identified as a target of circ_ROBO2, and circ_ROBO2 knockdown attenuated ox-LDL-induced damage in CMECs by sponging miR-186-5p. Tripartite motif containing 14 (TRIM14) acted as a target of miR-186-5p, and TRIM14 overexpression alleviated miR-186-5p-mediated inhibitory effect on ox-LDL-induced injury in CMECs. Circ_ROBO2 positively regulated TRIM14 expression by sponging miR-186-5p.
Conclusion:
Circ_ROBO2 played a promoting role in ox-LDL-induced CMECs injury by sponging miR-186-5p and regulating TRIM14, providing a promising treatment strategy for CAD.
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