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Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation
Published on: May 31, 2016
Ox-LDL-Induced Vascular Smooth Muscle Cell Dysfunction Partly Depends on the Circ_0044073/miR-377-3p/AURKA Axis in
Qiang Wang1, Tao Wang1, Siyuan Liang1
1Department of Vascular Surgery, Taizhou University Affiliated Municipal Hospital.
Insights
Circular RNA hsa_circ_0044073 (circ_0044073) promotes atherosclerosis by sponging miR-377-3p, leading to increased AURKA expression. Inhibiting circ_0044073 may offer a therapeutic strategy for atherosclerosis.
Area of Science:
- Cardiovascular Biology
- Molecular Biology
- RNA Biology
Background:
- Atherosclerosis (AS) is a primary cause of cardiovascular diseases.
- Circular RNA hsa_circ_0044073 (circ_0044073) is implicated in AS progression, but its regulatory mechanism is not fully understood.
Purpose of the Study:
- To elucidate the specific regulatory mechanism of circ_0044073 in AS progression.
- To investigate the role of circ_0044073 in oxidized low-density lipoprotein (Ox-LDL)-induced human vascular smooth muscle cell (VSMC) dysfunction.
Main Methods:
- Utilized Ox-LDL-stimulated human VSMCs as an AS cell model.
- Assessed circ_0044073 expression via RT-qPCR.
- Evaluated cell viability, proliferation, migration, and invasion using MTT, EDU, colony formation, and Transwell assays.
- Determined protein levels via Western blotting.
- Validated regulatory mechanisms using bioinformatics, dual-luciferase reporter, and RNA pull-down assays.
Main Results:
- Circ_0044073 expression was significantly elevated in AS patient serum and Ox-LDL-stimulated VSMCs.
- Circ_0044073 acts as a sponge for miR-377-3p.
- Knockdown of circ_0044073 or overexpression of miR-377-3p attenuated Ox-LDL-induced VSMC proliferation, migration, invasion, and inflammation.
- AURKA was identified as a direct target of miR-377-3p, and circ_0044073 regulated AURKA expression via miR-377-3p adsorption.
- Overexpression of AURKA partially reversed the inhibitory effects of circ_0044073 inhibition.
Conclusions:
- Circ_0044073 promotes AS progression by upregulating AURKA expression through sponging miR-377-3p.
- Circ_0044073 represents a potential therapeutic target for atherosclerosis.
Abstract:
Atherosclerosis (AS) is the main reason for most cardiovascular diseases. Circular RNA hsa_circ_0044073 (circ_0044073) has been found to promote AS progression. However, the specific regulatory mechanism of circ_0044073 in AS progression remains unclear.In this study, oxidized low-density lipoprotein (Ox-LDL) -stimulated human vascular smooth muscle cells (VSMCs) were used as AS cell models. The expression changes of circ_0044073 in serum samples and Ox-LDL-stimulated human VSMCs were assessed via real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability, proliferation, colony formation, migration, and invasion were assessed using 3- (4,5-Dimethylthiazol-2-yl) -2,5-Diphenyltetrazolium Bromide (MTT), 5-ethynyl-2'-deoxyuridine (EDU), colony formation, and transwell assays. Some protein levels were detected via Western blotting. The regulatory mechanism of circ_0044073 was predicted using bioinformatics analysis and validated by dual-luciferase reporter and RNA pull-down assays.We observed an overt increase in circ_0044073 expression in serum samples derived from AS patients and Ox-LDL-stimulated human VSMCs. Circ_0044073 was identified as a miR-377-3p sponge. Either circ_0044073 knockdown or miR-377-3p overexpression could impair Ox-LDL-induced human VSMC proliferation, migration, invasion, and inflammation. AURKA served as a miR-377-3p target, and circ_0044073 regulated AURKA expression by adsorbing miR-377-3p. Furthermore, AURKA overexpression partly reversed the effects of circ_0044073 inhibition on Ox-LDL-induced human VSMC proliferation, migration, invasion, and inflammation.Circ_0044073 promoted AS progression by elevating AURKA expression by functioning as a miR-377-3p sponge. Providing a proof-of-concept demonstration to support circ_0044073 might be a target for AS treatment.
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