Zhaohong Kong1,2, Yunfeng Wang1,2, Yudi Zhang1,2, Wei Shan1,2,3,4, Jianping Wu2,3, Qun Wang1,2,4. 1. Department of Neurology, Beijing Tiantan Hospital, Capital Medical University, Beijing, China. 2. China National Clinical Research Center for Neurological Diseases, Beijing, China. 3. Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing, China. 4. Beijing Institute for Brain Disorders, Beijing, China.
Abstract
BACKGROUND: Effective regulation of the biological function of endothelial progenitor cells (EPCs) is of great importance in its clinical application. This study aimed to explore the effect of microRNA-126 (miR-126) on the proliferation and migration of EPCs and the possible mechanism involved. METHODS: EPCs was isolated and cultured in vitro, and differences in the expression of miR-126 in endothelial cells (ECs) and EPCs, respectively, were detected by quantitative real-time PCR (RT-PCR). EPCs proliferation was then observed through CCK8 and colony formation experiments. Flow cytometry was also used to observe changes in the cycle and apoptosis of EPCs, and their migration ability was detected by scratch healing and Transwell assays. RT-PCR and Western blotting were carried out to observe the expression of key mRNA molecules and proteins of the Notch pathway. RESULTS: The relative expression of miR-126 in the EPCs group were 1.91±0.21, which was significantly higher than that in the EC group (1.25±0.06, P<0.05). When si-miR-126 and si-NC were transfected into the EPCs, it was found that the proliferation ability of cells in the si-miR-126 group decreased significantly (P<0.05), the apoptotic rate of the cells transfected with si-miR-126 was significantly increased, and the cell cycle was blocked at G0/G1 phase. RT-PCR and Western blotting demonstrated that the mRNA and protein expressions of Notch 1 and HES were significantly decreased in the si-miR-126 group. CONCLUSIONS: miR-126 can effectively promote the proliferation, invasion, and migration of EPCS, while inhibiting apoptosis, through the Notch1 pathway. 2020 Cardiovascular Diagnosis and Therapy. All rights reserved.
BACKGROUND: Effective regulation of the biological function of endothelial progenitor cells (EPCs) is of great importance in its clinical application. This study aimed to explore the effect of microRNA-126 (miR-126) on the proliferation and migration of EPCs and the possible mechanism involved. METHODS: EPCs was isolated and cultured in vitro, and differences in the expression of miR-126 in endothelial cells (ECs) and EPCs, respectively, were detected by quantitative real-time PCR (RT-PCR). EPCs proliferation was then observed through CCK8 and colony formation experiments. Flow cytometry was also used to observe changes in the cycle and apoptosis of EPCs, and their migration ability was detected by scratch healing and Transwell assays. RT-PCR and Western blotting were carried out to observe the expression of key mRNA molecules and proteins of the Notch pathway. RESULTS: The relative expression of miR-126 in the EPCs group were 1.91±0.21, which was significantly higher than that in the EC group (1.25±0.06, P<0.05). When si-miR-126 and si-NC were transfected into the EPCs, it was found that the proliferation ability of cells in the si-miR-126 group decreased significantly (P<0.05), the apoptotic rate of the cells transfected with si-miR-126 was significantly increased, and the cell cycle was blocked at G0/G1 phase. RT-PCR and Western blotting demonstrated that the mRNA and protein expressions of Notch 1 and HES were significantly decreased in the si-miR-126 group. CONCLUSIONS: miR-126 can effectively promote the proliferation, invasion, and migration of EPCS, while inhibiting apoptosis, through the Notch1 pathway. 2020 Cardiovascular Diagnosis and Therapy. All rights reserved.
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