Inhibition of microRNA-218 reduces HIF-1α by targeting on Robo1 in mice aortic endothelial cells under intermittent hypoxia
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Kai-Xiong Liu1,2,3, Qin Chen4, Gong-Ping Chen1,2,3, Jian-Chai Huang1,2,3, Jie-Feng Huang1,2,3, Xin-Ru He1,2,3, Ting Lin1,2,3 and Qi-Chang Lin1,2,3
1Department of Respiratory Disease, The First Affiliated Hospital, Fujian Medical University, China
2Laboratory of Respiratory Disease of Fujian Medical University, Fuzhou, China
3Fujian Provincial Sleep-disordered Breathing Clinic Center, Fuzhou, China
4Integrated Chinese and Western Medicine Colleges, Fujian University of Traditional Chinese Medicine, Fuzhou, China
Qi-Chang Lin, email: [email protected]
Keywords: microRNA-218; intermittent hypoxia-inducible factor-1α; apoptosis; normal mice aortic endothelial cells
Received: August 18, 2017 Accepted: September 22, 2017 Published: October 26, 2017
Objective: To investigate the effects of miR-218 on expression of hypoxia-inducible factors 1α (HIF-1α), vascular endothelial growth factor (VEGF) and cell apoptosis in normal mice aortic endothelial cells under intermittent hypoxia (IH) condition. Methods: Anti-miR-218 inhibitor, miR-negative control and miR-218 mimic were used to tranfect the cells in different groups under IH condition. Both RT-PCR and Western blot were used to determine the expressions of HIF-1α and VEGF. Akt, p-Akt and cell apoptosis related proteins bcl-2, bax and caspase-3 and roundabout 1 (Robo1) were measured using Western blot. Cell apoptosis was evaluated by flow cytometry. Statistical analysis was performed using SPSS 18.0. Results: Expression of miR-218 was significantly up-regulated in the IH group and was significantly inhibited when cells were transfected with miR-218 inhibitor. Down regulation of miR-218 could reduce the expression of HIF-1α and VEGF under intermittent hypoxia condition. In cells transfected with miR-218 mimic, expression of HIF-1α and VEGF significantly increased compared with the control. However, when treated with LY294002, the expression of HIF-1α and VEGF both decreased. Apoptosis assay showed that down regulation of miR-218 could inhibit intermittent hypoxia induced cell apoptosis, decrease expression of caspase-3 and bax and increase expression of bcl-2 under intermittent hypoxia condition. At last, silencing Robo1 could significantly enhance the expression of HIF-1α under IH condition. Conclusion: Inhibition of miR-218 could reduce the expression of HIF-1α and protect against IH-induced apoptosis in mice aortic endothelial cells. The effects were associated with PI3K/AKT pathway and might through targeting of Robo1.
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