Inhibition of microRNA-218 reduces HIF-1α by targeting on Robo1 in mice aortic endothelial cells under intermittent hypoxia.
Inhibition of microRNA-218 reduces HIF-1α by targeting on Robo1 in mice aortic endothelial cells under intermittent hypoxia.
复制标题
DOI：
10.18632/oncotarget.22239
复制
发表时间：
2017-11-28
期刊：
影响因子：
--
通讯作者：
Lin QC
中科院分区：
文献类型：
--
作者：
Liu KX;Chen Q;Chen GP;Huang JC;Huang JF;He XR;Lin T;Lin QC
目的：研究miR218对间歇性低氧(IH)状态下正常小鼠主动脉内皮细胞缺氧诱导因子1α(HIF1α)、血管内皮生长因子(VEGFR)表达及细胞凋亡的影响。在IH条件下，用抗miR-218抑制剂、miR阴性对照和miR-218模拟物对不同组的细胞进行转导。RT-PCR法和Western印迹法检测缺氧诱导因子-1、α和血管内皮生长因子的表达。用Western印迹法检测AKT、p-Akt和细胞凋亡相关蛋白bcl2、bax、caspase-3和转盘1(Robo1)的表达。流式细胞仪检测细胞凋亡率。采用SPSS18.0统计软件进行统计分析。在IH组，miR-218的表达显著上调，而当miR-218抑制剂转染细胞时，miR-218的表达被显著抑制。在间歇性低氧条件下，下调miR218可减少缺氧诱导因子-1α和血管内皮生长因子的表达。与对照组相比，转染miR218模拟物的细胞中，HIF1α和血管内皮生长因子的表达显著增加。而经LY294002处理后，缺氧诱导因子-1α和血管内皮生长因子的表达均降低。细胞凋亡实验显示，在间歇性低氧条件下，miR-218的下调可抑制间歇性低氧诱导的细胞凋亡，降低caspase-3和bax的表达，增加bcl2的表达。最后，在IH条件下，沉默Robo1可显著增强HIF-1α的表达。抑制miR218可降低缺氧诱导因子-1α的表达，对IH诱导的小鼠血管内皮细胞凋亡具有保护作用。这种作用可能与PI3K/AKT途径有关，并可能通过靶向Robo1发挥作用。
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. 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. 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. 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.
登录
查看更多内容
影响因子：
5.6
作者：
Kanichai, Manoj;Ferguson, Damien;Campbell, Veronica A.
通讯作者：
Campbell, Veronica A.
影响因子：
4.6
作者：
Fish, Jason E.;Wythe, Joshua D.;Woo, Stephanie
通讯作者：
Woo, Stephanie
microRNA-494 的过表达通过 PI3K/Akt 途径上调缺氧诱导因子 1 α 表达，并防止缺氧诱导的细胞凋亡
影响因子：
11
作者：
Sun G;Zhou Y;Li H;Guo Y;Shan J;Xia M;Li Y;Li S;Long D;Feng L
通讯作者：
Feng L
影响因子：
3.4
作者：
Wang Z;Si LY
通讯作者：
Si LY
影响因子：
2.3
作者：
Nanduri, Jayasri;Yuan, Guoxiang;Kumar, Ganesh K.;Semenza, Gregg L.;Prabhakar, Nanduri R.
通讯作者：
Prabhakar, Nanduri R.