H Chang1, Z-B Li, J-Y Wu, L Zhang. 1. Department of Cardiovascular Medicine, China-Japan Union Hospital, Jilin University, Changchun, Jilin, China. Zhanglei99@jlu.edu.cn.
Abstract
OBJECTIVE: To investigate the effect of circular RNA circ-100338 on angiogenesis of human umbilical vein endothelial cells (HUVEC) cells after hypoxia/reoxygenation (H/R) and its molecular mechanism. MATERIALS AND METHODS: We evaluated the role of circ-100338 in coronary artery endothelial cells using human coronary endothelial cells (HCAEC). Then, we verified the function of circ-100338 in HUVEC cells through cell counting kit-8 (CCK8), scratch test, Tube forming experiment, 5-Ethynyl-2'-deoxyuridine (EdU) staining. Dual-Luciferase reporter gene experiment and RNA Pull-Down experiments were used to detect the binding effect of circ-100338 and miR-200a-3p, miR-200a-3p and FUS. RESULTS: QRT-PCR results showed that the expression of circ-100338 decreased in HCAEC after H/R treatment. Overexpression of circ-100338 promotes angiogenesis. The Dual-Luciferase reporter gene assay and RNA pull-down assay consistently indicated the specific binding effect between circ-100338 and miR-200a-3p, miR-200a-3p and FUS, and circ-100338 promoted the angiogenesis phenotype in HUVEC cells. CONCLUSIONS: CircRNA-100338 may inhibit the function of miRNA-200a-3p by combining with miRNA-200a-3p, and then miRNA-200a-3p plays a role in regulating FUS, thereby regulating the state of angiogenesis.
OBJECTIVE: To investigate the effect of circular RNA circ-100338 on angiogenesis of human umbilical vein endothelial cells (HUVEC) cells after hypoxia/reoxygenation (H/R) and its molecular mechanism. MATERIALS AND METHODS: We evaluated the role of circ-100338 in coronary artery endothelial cells using human coronary endothelial cells (HCAEC). Then, we verified the function of circ-100338 in HUVEC cells through cell counting kit-8 (CCK8), scratch test, Tube forming experiment, 5-Ethynyl-2'-deoxyuridine (EdU) staining. Dual-Luciferase reporter gene experiment and RNA Pull-Down experiments were used to detect the binding effect of circ-100338 and miR-200a-3p, miR-200a-3p and FUS. RESULTS: QRT-PCR results showed that the expression of circ-100338 decreased in HCAEC after H/R treatment. Overexpression of circ-100338 promotes angiogenesis. The Dual-Luciferase reporter gene assay and RNA pull-down assay consistently indicated the specific binding effect between circ-100338 and miR-200a-3p, miR-200a-3p and FUS, and circ-100338 promoted the angiogenesis phenotype in HUVEC cells. CONCLUSIONS:CircRNA-100338 may inhibit the function of miRNA-200a-3p by combining with miRNA-200a-3p, and then miRNA-200a-3p plays a role in regulating FUS, thereby regulating the state of angiogenesis.