miR-126 regulates platelet-derived growth factor receptor-α expression and migration of primary human osteoblasts
miR-126 regulates platelet-derived growth factor receptor-α expression and migration of primary human osteoblasts
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DOI：
10.1515/hsz-2014-0168
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发表时间：
2015-01-01
影响因子：
3.7
通讯作者：
Finkenzeller, Guenter
中科院分区：
文献类型：
--
作者：
Schmidt, Yvonne;Simunovic, Filip;Finkenzeller, Guenter
充足的血管化是骨发育、骨折愈合和骨组织工程的基本要求。我们先前已经描述了原代人成骨细胞（hOBs）和人内皮细胞（HUVECs）的共培养，旨在研究这些细胞之间的相互作用。在这个系统中，我们表明，这两种细胞类型的共培养导致hOB中血小板衍生生长因子受体-α（PDGFR-α）的下调，这是mRNA稳定性降低的结果。在目前的研究中，我们研究了microRNA在这一过程中的可能参与。首先，我们对与HUVEC共培养后的成骨细胞miRNAs进行了微阵列分析，揭示了miR-126的上调。这一结果通过RT-qPCR得到证实，我们观察到这种增加依赖于直接的细胞间接触。功能获得和功能丧失实验表明，miR-126是PDGFR-α mRNA的负调控因子。此外，hOB的迁移受到miR-126过表达的抑制，并受到miR-126抑制的刺激。向hOB培养物中添加PDGFR-α阻断抗体也抑制hOB迁移。miR-126调节对成骨细胞增殖、凋亡率或分化没有影响。总之，我们报告了miR-126/PDGFR-α系统调节人成骨细胞的迁移行为，而不影响细胞的存活和分化。
Adequate vascularization is an essential requirement for bone development, fracture healing and bone tissue engineering. We have previously described the coculture of primary human osteoblasts (hOBs) and human endothelial cells (HUVECs), designed to investigate the interactions between these cells. In this system, we showed that cocultivation of these two cell types leads to a downregulation of platelet-derived growth factor receptor-alpha (PDGFR-alpha) in hOBs, which was a consequence of reduced mRNA stability. In the current study we investigated the possible involvement of microRNAs in this process. Firstly, we performed a microarray analysis of osteoblastic miRNAs following cocultivation with HUVECs, revealing an upregulation of miR-126. This result was confirmed by RT-qPCR, and we observed that the increase is dependent on direct cell-to-cell contacts. Gain-of-function and loss-of-function experiments showed that miR-126 is a negative regulator of PDGFR-alpha mRNA. Additionally, migration of hOBs was inhibited by miR-126 overexpression and stimulated by miR-126 inhibition. Addition of PDGFR-alpha blocking antibody to hOB culture also inhibited hOB migration. There was no effect of miR-126 modulation on osteoblast proliferation, apoptosis rate or differentiation. In conclusion, we report that the miR-126/PDGFR-alpha system regulates the migratory behavior of human osteoblasts, without exerting effects on cell survival and differentiation.