MicroRNA-130a regulates autophagy of endothelial progenitor cells through Runx3
MicroRNA-130a regulates autophagy of endothelial progenitor cells through Runx3
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MicroRNA-130a通过Runx3调节内皮祖细胞自噬
DOI：
10.1111/1440-1681.12227
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发表时间：
2014-05-01
影响因子：
2.9
通讯作者：
Li, Yi-Gang
中科院分区：
文献类型：
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作者：
Xu, Quanfu;Meng, Shu;Li, Yi-Gang
内皮祖细胞功能障碍与糖尿病血管疾病有关。MicroRNAs （miRNAs）是多种细胞过程的关键调控因子，包括血管生成。我们最近报道，miR-130a在2型糖尿病（DM）患者中下调可导致EPC功能障碍，包括细胞凋亡增加，可能是通过其靶矮子相关转录因子3 （Runx3）。然而，miR-130a是否影响EPC的自噬尚不清楚。本研究的目的是探讨miR-130a对EPC自噬和细胞死亡的影响，以及其Beclin 1 （BECN1，自噬体形成的启动物）和抗凋亡蛋白Bcl2 （BECN1结合并失活）的表达，以及Runx3在介导这些作用中的作用。从糖尿病患者和非糖尿病对照组外周血单个核细胞中培养EPC。用miR-130a抑制剂或mimic-miR-130a或mimic-miR-130a加表达Runx3的慢病毒载体转染细胞，以操纵miR-130a和/或Runx3水平。透射电镜观察自噬体数量，流式细胞术观察细胞死亡情况。实时聚合酶链反应检测Beclin1 mRNA表达量，western blotting检测Beclin1和Bcl2蛋白表达量。在DM患者的EPC中，自噬体的数量和Beclin1的表达均增加。抑制miR-130a增加了自噬体的数量和Beclin1的表达，但减弱了Bcl2的表达。过表达miR-130a可降低自噬体数量、细胞死亡和Beclin1表达，但可促进Bcl2表达；这些作用是由Runx3介导的。综上所述，miR-130a通过Runx3调控Bcl-2和Beclin1的表达，在维持正常自噬水平和促进EPC存活方面发挥重要作用。MiR-130a可能是连接EPC细胞凋亡和自噬的调节因子。
Dysfunction of endothelial progenitor cells (EPC) contribute to diabetic vascular disease. MicroRNAs (miRNAs) are key regulators of diverse cellular processes, including angiogenesis. We recently reported that downregulated miR-130a in patients with Type 2 diabetes mellitus (DM) results in EPC dysfunction, including increased apoptosis, likely via its target runt-related transcription factor 3 (Runx3). However, whether miR-130a affects the autophagy of EPC is unknown. The aim of the present study was to explore the effects of miR-130a on the autophagy and cell death of EPC, as well as their expression of Beclin 1 (BECN1; an initiator of autophagosome formation) and the anti-apoptotic protein Bcl2 (which binds to and inactivates BECN1), and the role of Runx3 in mediating these effects. The EPC were cultured from peripheral blood mononuclear cells of diabetic patients and non-diabetic controls. Cells were transfected with an miR-130a inhibitor, or mimic-miR-130a or mimic-miR-130a plus lentiviral vector expressing Runx3 to manipulate miR-130a and/or Runx3 levels. The number of autophagosomes was counted under transmission electron microscopy and cell death was examined by flow cytometry. The mRNA expression of Beclin1 was measured by real-time polymerase chain reaction and the protein expression of Beclin1 and Bcl2 was determined by western blotting. Both the number of autophagosomes and Beclin1 expression were increased in EPC from patients with DM. Inhibition of miR-130a increased the number of autophagosomes and Beclin1 expression, but attenuated Bcl2 expression. Overexpression of miR-130a decreased the number of autophagosomes, cell death and Beclin1 expression, but promoted Bcl2 expression; these effects were mediated by Runx3. In conclusion, miR-130a is important for maintaining normal autophagy levels and promoting the survival of EPC via regulation of Bcl-2 and Beclin1 expression, via Runx3. MiR-130a may be a regulator linking apoptosis and the autophagy of EPC.