Exosomal miR-130a-3p regulates osteogenic differentiation of Human Adipose-Derived stem cells through mediating SIRT7/Wnt/β-catenin axis.
Exosomal miR-130a-3p regulates osteogenic differentiation of Human Adipose-Derived stem cells through mediating SIRT7/Wnt/β-catenin axis.
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外泌体 miR-130a-3p 通过介导 SIRT7/Wnt/β-catenin 轴调节人脂肪干细胞的成骨分化
DOI：
10.1111/cpr.12890
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发表时间：
2020-10
影响因子：
8.5
通讯作者：
Sun X
中科院分区：
文献类型：
--
作者：
Yang S;Guo S;Tong S;Sun X
我们发现外泌体可能促进脂肪源性干细胞（ADSC）成骨分化，阐明其具体分子机制对临床骨再生具有深远意义。通过慢病毒转染的方式，构建了miR-130a-3p过表达和敲低的ADSCs。采用茜素红S检测钙沉积，采用qPCR检测成骨相关基因，验证miR-130a-3p对ADSCs成骨分化的影响。采用CCK-8检测miR-130a-3p对ADSC增殖的影响。通过双荧光素酶报告基因测定验证了 miR-130a-3p 和 SIRT7 之间的靶结合。此外，通过检测成骨相关基因和蛋白以及碱性磷酸酶活性，进一步验证了Wnt信号通路在miR-130a-3p调控ADSCs成骨和分化中的作用。 (a) miR-130a-3p 的过表达可以增强 ADSC 的成骨分化，同时降低 miR-130a-3p 的靶标 SIRT7 的蛋白和 mRNA 水平。 (b)我们的研究进一步发现，miR-130a-3p的过度表达导致SIRT7表达下调，同时Wnt信号通路相关蛋白上调。 (c) miR-130a-3p 的过表达抑制 ADSC 的增殖，而敲低则促进其增殖。研究结果表明，外泌体miR-130a-3p可以部分通过介导SIRT7/Wnt/β-catenin轴来促进ADSCs的成骨分化，从而促进外泌体microRNA在骨再生领域的应用。为了促进外泌体microRNA在骨再生领域的应用，我们在此研究外泌体miR-130a-3p是否可以部分通过介导SIRT7/Wnt/β-catenin轴来促进ADSCs的成骨分化。当外泌体与细胞膜融合时，“货物”就会被释放。外泌体中富集的 miR-130a-3p 与 SIRT7 mRNA 结合，导致 SIRT7 mRNA 降解。最终，Wnt信号通路从“关闭状态”变为“开启状态”，启动ADSCs的成骨分化过程。
It is of profound significance for clinical bone regeneration to clarify the specific molecular mechanism from which we found that osteogenic differentiation of adipose‐derived stem cells (ADSCs) will be probably promoted by exosomes. By means of lentiviral transfection, miR‐130a‐3p overexpression and knockdown ADSCs were constructed. Alizarin Red S was used to detect the calcium deposits, and qPCR was used to detect osteogenesis‐related genes, to verify the effect of miR‐130a‐3p on the osteogenic differentiation of ADSCs. CCK‐8 was used to detect the effect of miR‐130a‐3p on the proliferation of ADSCs. The target binding between miR‐130a‐3p and SIRT7 was verified by dual‐luciferase reporter gene assay. Furthermore, the role of Wnt signalling pathway in the regulation of ADSCs osteogenesis and differentiation by miR‐130a‐3p was further verified by detecting osteogenic‐related genes and proteins and alkaline phosphatase activity. (a) Overexpression of miR‐130a‐3p can enhance the osteogenic differentiation of ADSCs while reducing protein and mRNA levels of SIRT7, a target of miR‐130a‐3p. (b) Our study further found that overexpression of miR‐130a‐3p leads to down‐regulation of SIRT7 expression with up‐regulation of Wnt signalling pathway‐associated protein. (c) Overexpression of miR‐130a‐3p inhibited proliferation of ADSCs, while knockdown promoted it. The obtained findings indicate that exosomal miR‐130a‐3p can promote osteogenic differentiation of ADSCs partly by mediating SIRT7/Wnt/β‐catenin axis, which will hence promote the application of exosomal microRNA in the field of bone regeneration. In order to promote the application of exosomal microRNA in the field of bone regeneration, we investigate herein whether exosomal miR‐130a‐3p can promote osteogenic differentiation of ADSCs partly by mediating SIRT7/Wnt/β‐catenin axis. When exosomes fuse with the cell membrane, the 'cargos' are released. Enriched miR‐130a‐3p in exosomes binds to SIRT7 mRNA, resulting in SIRT7 mRNA degradation. Eventually, Wnt signaling pathway changes from 'off state' to 'on state', initiating the osteogenic differentiation process of ADSCs.
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影响因子：
9.2
作者：
Ding Y;Yang H;Wang Y;Chen J;Ji Z;Sun H
通讯作者：
Sun H
影响因子：
4.8
作者：
Cohen-Kfir, Einav;Artsi, Hanna;Dresner-Pollak, Rivka
通讯作者：
Dresner-Pollak, Rivka
影响因子：
2.3
作者：
Waki T;Lee SY;Niikura T;Iwakura T;Dogaki Y;Okumachi E;Oe K;Kuroda R;Kurosaka M
通讯作者：
Kurosaka M
影响因子：
4.8
作者：
Fang, Shanhong;Li, Yongfeng;Chen, Peng
通讯作者：
Chen, Peng
影响因子：
13.5
作者：
Kim, Jeong Kyu;Noh, Ji Heon;Nam, Suk Woo
通讯作者：
Nam, Suk Woo