MiR-590-3p regulates osteogenic differentiation of human mesenchymal stem cells by regulating APC gene
MiR-590-3p regulates osteogenic differentiation of human mesenchymal stem cells by regulating APC gene
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DOI：
10.1016/j.bbrc.2016.08.160
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发表时间：
2016-09-30
影响因子：
3.1
通讯作者：
Zhou, Lei
中科院分区：
文献类型：
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作者：
Wu, Siyuan;Liu, Weizhen;Zhou, Lei
本研究旨在探讨miR-590- 3 p在人骨髓间充质干细胞（hMSC）成骨分化中的作用。培养HMSC，转染空载体、miR-590- 3 p载体，构建对照hMSC和miR-590- 3 p过表达hMSC。用寡核苷酸瞬时转染产生miR-590- 3 p抑制的hMSC。分析所有3组细胞的成骨水平以及Wnt/β-连环蛋白信号通路活性。茜素红染色结果表明，miR-590- 3 p的过表达可显著促进矿化沉积。成骨标志物ALP、OC和SOST也上调。双荧光素酶和western blot分析结果表明，miR-590- 3 p选择性结合APC mRNA的3 'UTR。过表达miR-590- 3 p可通过抑制APC的mRNA水平，稳定β-catenin，提高下游靶基因的转录活性。这些结果表明miR-590- 3 p可以通过抑制APC表达和稳定β-catenin来促进成骨分化。(C)2016 Elsevier Inc. All rights reserved.
The present study aimed to investigate miR-590-3p's role in osteogenic differentiation of human mesenchymal stem cells (hMSC). HMSC were cultured and transfected with empty vector, miR-590-3p vector to construct control hMSC and miR-590-3p overexpression hMSC. MiR-590-3p suppressed hMSC was created with oligonucleotide transient transfection. All 3 groups of cells were analyzed for their osteogenic level as well as Wnt/beta-catenin signaling pathway activities. The results of alizarin red staining indicated that overexpression of miR-590-3p significantly enhanced mineralized deposition. Osteogenic markers including ALP, OC and SOST were also up-regulated. The result of dual-luciferase and western blot analysis indicated that miR-590-3p bound to 3'UTR of APC mRNA selectively. By suppressing mRNA level of APC, over-expressed miR-590-3p stabilized beta-catenin and increased the transcription activities of downstream target genes. These result suggested that miR-590-3p can promote osteogenic differentiation via suppressing APC expression and stabilizing beta-catenin. (C) 2016 Elsevier Inc. All rights reserved.