Kong Zhaohong, Wang Yunfeng, Zhang Yudi, Shan Wei, Wu Jianping, Wang Qun
Department of Neurology, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.
China National Clinical Research Center for Neurological Diseases, Beijing, China.
Cardiovasc Diagn Ther. 2020 Jun;10(3):490-499. doi: 10.21037/cdt-20-178.
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.
EPCs was isolated and cultured , 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.
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.
miR-126 can effectively promote the proliferation, invasion, and migration of EPCS, while inhibiting apoptosis, through the Notch1 pathway.
有效调控内皮祖细胞（EPCs）的生物学功能对其临床应用至关重要。本研究旨在探讨微小RNA-126（miR-126）对EPCs增殖和迁移的影响及其可能的作用机制。
分离培养EPCs，通过定量实时聚合酶链反应（RT-PCR）检测内皮细胞（ECs）和EPCs中miR-126的表达差异。然后通过CCK8和集落形成实验观察EPCs的增殖情况。采用流式细胞术观察EPCs的细胞周期和凋亡变化，通过划痕愈合实验和Transwell实验检测其迁移能力。进行RT-PCR和蛋白质印迹法观察Notch信号通路关键mRNA分子和蛋白质的表达。
EPCs组中miR-126的相对表达量为1.91±0.21，显著高于EC组（1.25±0.06，P<0.05）。将si-miR-126和si-NC转染至EPCs后，发现si-miR-126组细胞的增殖能力显著降低（P<0.05），转染si-miR-126的细胞凋亡率显著升高，细胞周期阻滞在G0/G1期。RT-PCR和蛋白质印迹法显示，si-miR-126组中Notch 1和HES的mRNA和蛋白质表达显著降低。
miR-126可通过Notch1信号通路有效促进EPCs的增殖、侵袭和迁移，同时抑制细胞凋亡。
