miR-144/451通过干扰14-3-3ζ/AKT/ROS正反馈通路抑制ROS的产生

miR-144/451 inhibits ROS production by suppressing 14-3-3ζ/AKT/ROS feedback loop

利用miR-144/451小分子RNA基因敲除小鼠分离骨髓有核红细胞,使用流式细胞术、定量PCR和Western blot等方法证明miR-144/451是否通过干扰14-3-3ζ/AKT/ROS正反馈通路抑制超氧化活性氧簇(ROS)的产生。结果表明:miR-144/451敲除小鼠呈现小细胞性溶血性贫血,可能是miR-144/451敲后除引起14-3-3ζ增高,而14-3-3ζ增高可引起轻度AKT磷酸化,从而引起Foxo3入核障碍;ROS水平增高,进而激活AKT磷酸化,且ROS促AKT磷酸化程度比14-3-3ζ更强,从而加强14-3-3ζ与Foxo3的结合,引起Foxo3进一步出核,形成正反馈调节机制;ROS水平进一步增高,导致红细胞凋亡加速,小鼠呈现贫血症状。

We isolated nucleated bone marrow erythroid cells from miR-144/451 knockout mice and validated our hypothesis that miR-144/451 may inhibit ROS production by suppressing 14-3-3ζ/AKT/ROS feedback loop using analyses such as flow cytometry, quantitative PCR and western blot. Here we discovered that miR-144/451 knockout mice exhibited microcytic anemia. The underlying mechanism was that loss of miR-144/451 gene locus increased the level of 14-3-3ζ which led to the phosphorylation of AKT and subsequent exclusion of transcription factor Foxo3 from nucleus and thus the increasing of ROS. High ROS level further activated AKT and this activation of AKT by ROS was even stronger than that by overexpressed 14-3-3ζ, This promoted further binding of 14-3-3ζ with and subsequent exclusion of Foxo3 from nucleus, forming a 14-3-3ζ/AKT/ROS positive feedback loop that led to increased ROS production and the death of erythrocytes, and thus the anemia.