MicroRNA-10a Influences Osteoblast Differentiation and Angiogenesis by Regulating β-Catenin Expression
MicroRNA-10a Influences Osteoblast Differentiation and Angiogenesis by Regulating β-Catenin Expression
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DOI：
10.1159/000438576
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发表时间：
2015-01-01
影响因子：
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通讯作者：
He, Xijing
中科院分区：
文献类型：
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作者：
Li, Jun;Zhang, Yongqing;He, Xijing
背景/目的：阐明成骨细胞分化和血管生成的分子机制对改善骨相关疾病的治疗具有重要意义。在本研究中，我们检测了microRNA (miR)-10a在小鼠脐静脉内皮细胞（MUVECs） MC3T3-E1细胞分化和促血管生成活性中的作用。方法：采用小鼠成骨前细胞系MC3T3-E1和MUVECs进行实验。转染miR-10a模拟物或抑制剂后，经或不经LiCI预处理，RT-PCR检测miR-10a、ALP、Runx2、Osx、OC和DIx5的表达。用BMP2培养MC3T3-E1细胞向骨细胞分化，用ALP和ARS染色检测MC3T3-E1细胞的成骨分化。采用MIT检测细胞活力，Western blotting检测13-catenin、LEF1、cyclinD1、MMP2、VEGF蛋白表达；ELISA法检测VEGF、VE-cadherin释放量，检测muvec的迁移及小管形成情况。结果：成骨分化过程中mir -10a表达明显下调。过表达miR-10a抑制了MC3T3-E1细胞的成骨分化，有效地降低了muvec的增殖、迁移、VEGF表达、VE-cadherin浓度和试管形成，而miR-10a沉默则增强了这些过程。进一步的机制分析表明，miR-10a过表达在蛋白和转录水平上降低了13-连环蛋白，而Wnt信号激活剂Lid预处理部分减弱了miR-10a过表达对成骨细胞分化和血管生成的抑制作用。结论：我们的研究结果表明，miR-10a通过调节13-catenin的表达，抑制MC3T3-E1细胞的成骨细胞分化和促进MUVECs的血管生成活性，是治疗骨再生相关疾病的一个新的潜在治疗靶点。(C) 2015作者：s . Karger AG，巴塞尔出版
Background/Aims: Elucidation of the molecular mechanisms governing osteoblast differentiation and angiogenesis are of great importance for improving the treatment of bone-related diseases. In this study, we examined the role of microRNA (miR)-10a in the differentiation of MC3T3-E1 cells and pro angiogenic activity of mouse umbilical vein endothelial cells (MUVECs). Methods: The murine pre-osteoblast cell line MC3T3-E1 and MUVECs were used in the experiment. After transfected with miR-10a mimics or inhibitors, with or without LiCI pretreatment, the miR-10a, ALP, Runx2, Osx, OC and DIx5 expression were assessed by RT-PCR. MC3T3-E1 cells were cultured with BMP2 to differentiate into bone cells, osteogenic differentiation of MC3T3-E1 cells were detected by ALP and ARS staining. Cell viability were analyzed by MIT and the protein expression of 13-catenin, LEF1, cyclinD1, MMP2, and VEGF were detected by Western blotting; VEGF and VE-cadherin release were assessed by ELISA, and the migration of MUVECs, as well as tube formation were also detected. Results: M iR-10a expression was obviously down-regulated during osteogenic differentiation. Overexpression of miR-10a inhibited osteogenic differentiation of MC3T3-E1 cells, effectively decreasing MUVECs proliferation, migration, VEGF expression, VE-cadherin concentrations, and tube formation in vitro, whereas miR-10a silence enhanced those processes. Further mechanism assays demonstrated that overexpression of miR-10a reduced the13-catenin at both protein and transcription level, while pretreatment with Wnt signaling activator Lid partially attenuated the suppression effects of miR-10a overexpression on osteoblast differentiation and angiogenesis. Conclusion: Our findings imply that miR-10a plays a suppressive role in osteoblast differentiation of MC3T3-E1 cells and pro angiogenic activity of MUVECs by regulating the 13-catenin expression, representing a novel and potential therapeutic target for the treatment of bone regeneration-related diseases. (C) 2015 The Author(s) Published by S. Karger AG, Basel