基因敲除
流式细胞术
下调和上调
细胞凋亡
免疫印迹
细胞生长
小RNA
内皮干细胞
细胞
基因沉默
分子生物学
化学
癌症研究
生物
体外
生物化学
基因
作者
Chunlei Chen,Jiguo Xu,Tianrun Huang,Qian Zhou
标识
DOI:10.1080/00207454.2023.2246100
摘要
Ischemic stroke (IS) is a highly prevalent type of stroke with very high rates of disability and death. As the regulatory role of circular RNAs (circRNAs) in various diseases has been revealed, we constructed a stroke cell model to analyze the action mechanism of hsa_circ_0005548 in IS.The abundance of hsa_circ_0005548, microRNA-362-3p (miR-362-3p) and E26 transformation specific-1 (ETS-1) were measured by real-time quantitative polymerase chain reaction (RT-qPCR) or western blot. We constructed an IS cell model in vitro by oxygen-glucose deprivation/reperfusion (OGD/R) treatment and analyzed cell proliferation, apoptosis and inflammatory response through the use of Cell Counting Kit-8 (CCK8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry and Enzyme-linked immunosorbent assay (ELISA), respectively. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were employed for the analysis of the relationship between miR-362-3p and hsa_circ_0005548 or ETS1.The higher abundance of hsa_circ_0005548 and ETS-1 and lower level of miR-362-3p were observed in human brain microvascular endothelial immortalized (HBMEC-IM) cells under OGD/R. Hsa_circ_0005548 downregulation mitigated OGD/R-induced HBMEC-IM cell injury. Mechanistically, hsa_circ_0005548 targeted miR-362-3p. MiR-362-3p knockdown reversed the effect of hsa_circ_0005548 silencing on OGD/R-induced HBMEC-IM cell injury. ETS1 was validated as a direct target of miR-362-3p, and miR-362-3p attenuated OGD/R-induced HBMEC-IM cell injury by ETS1. Moreover, hsa_circ_0005548 modulated ETS1 via miR-362-3p.Hsa_circ_0005548 knockdown repairs OGD/R-induced HBMEC-IM cell damage via miR-362-3p/ETS1 axis.
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