Abstract
BackgroundCircular RNAs have shown important regulatory roles in cardiovascular diseases, containing atherosclerosis (AS). We intended to explore the role of circ_0004104 in AS using oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cells and its associated mechanism.MethodsReal-time quantitative polymerase chain reaction and Western blot assay were conducted to analyze RNA levels and protein levels, respectively. Cell viability, apoptosis, angiogenic ability and inflammatory response were assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, flow cytometry, capillary-like network formation assay and enzyme-linked immunosorbent assay, respectively. Cell oxidative stress was assessed using commercial kits. Dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA-pull down assay were performed to verify the intermolecular interaction.Resultsox-LDL exposure up-regulated the level of circ_0004104 in HUVECs. ox-LDL exposure suppressed cell viability and angiogenic ability whereas promoted the apoptosis, inflammation and oxidative stress of HUVECs partly through up-regulating circ_0004104. MicroRNA-328-3p (miR-328-3p) was confirmed as a target of circ_0004104. MiR-328-3p interference largely reversed circ_0004104 silencing-mediated effects in HUVECs upon ox-LDL exposure. MiR-328-3p interacted with the 3′ untranslated region of tripartite motif 14, and circ_0004104 positively regulated TRIM14 expression by sponging miR-328-3p. TRIM14 overexpression largely overturned miR-328-3p accumulation-induced influences in HUVECs upon ox-LDL exposure.ConclusionCirc_0004104 knockdown attenuated ox-LDL-induced dysfunction in HUVECs via miR-328-3p-mediated regulation of TRIM14.
Highlights
Circular RNAs have shown important regulatory roles in cardiovascular diseases, containing atherosclerosis (AS)
Relative abundance of circ_0004104, SPARC and Tripartite motif 14 (TRIM14) was analyzed using the 2−ΔΔCt method with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as reference, while the fold change of miR328-3p was calculated using the 2−ΔΔCt method with U6 as reference
Characteristics of circ_0004104 in Human umbilical vein endothelial cells (HUVECs) Among several AS progression-associated circRNAs, including circ_0004104 [10], circ_0001879 [10], circ_0001445 [22], circ_0001599 [23], circ_0010283 [24] and circ_0007478 [25], we selected circ_0004104 for further analysis because it was the most significantly upregulated by oxidized low-density lipoprotein (ox-LDL) (100 μg/mL, 24 h) in HUVECs (Additional file 1: Figure 1 and Fig. 1a)
Summary
Circular RNAs have shown important regulatory roles in cardiovascular diseases, containing atherosclerosis (AS). We intended to explore the role of circ_0004104 in AS using oxidized low-density lipoprotein (ox-LDL)induced vascular endothelial cells and its associated mechanism. Many types of cells are involved in the pathogenetic process of atherosclerosis (AS), containing vascular endothelial cells and smooth muscle cells [1]. CircRNAs have been demonstrated to modulate the pathological process of human malignancies [6, 7]. Accumulating studies have identified the important roles of circRNAs in AS. Liu et al demonstrated that circ_0003204 suppressed proliferation and angiogenesis of ox-LDL-treated HUVECs [9]. The role and mechanism of circ_0004104 in AS progression remain largely unknown
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
