Luo Tao, Yang Xueqin, Sun Yan, Huang Xinqi, Zou Ling, Liu Jun
State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, Department of Endodontics, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
Department of Stomatology, Sichuan Cancer Hospital and Institute, Sichuan Cancer Center, School of Medicine, University of Electronic Science and Technology of China, Chengdu, China.
Cells Tissues Organs. 2019;208(3-4):148-157. doi: 10.1159/000506304. Epub 2020 Feb 25.
Osteogenic differentiation of human adipose tissue-derived stem cells (hASCs) is a complex process that is regulated by multiple factors, including microRNAs (miRNAs). The miRNA miR-20a was shown to promote bone formation from bone marrow-derived mesenchymal stem cells. However, the role of miR-20a in osteogenic differentiation of hASCs remains unclear. In this study, we systematically evaluated the function of miR-20a in regulating hASC osteogenesis in vitro. hASCs were transduced with miR-20a-overexpressing and miR-20a-sponge lentiviral vectors, with green fluorescent protein (GFP) as a control. The results showed that miR-20a transcription was upregulated after hASC mineralization. Compared with the miR-20a-sponge, GFP, and hASC groups, the miR-20a-overexpressing group showed higher alkaline phosphatase (ALP) activity on days 7 and 14. Moreover, the mRNA level of ALP increased significantly in the miR-20a-overexpressing group on day 14. Furthermore, the protein of the target gene PPARγ was decreased, and the osteogenic differentiation-associated proteins ALP, osteocalcin, and RUNX2 were upregulated. hASCs anchored to HA/β-TCP revealed a healthy polygonal morphology and developed cytoplasmic extensions. miR-20a promoted osteogenic differentiation of the cell scaffold. Taken together, these data -confirm that miRNA-20a promotes the osteogenesis of hASCs in vitro, and its essential role in vivo needs further -investigation.
人脂肪组织来源干细胞（hASCs）的成骨分化是一个受多种因素调控的复杂过程，这些因素包括微小RNA（miRNAs）。已表明微小RNA miR-20a可促进骨髓间充质干细胞的骨形成。然而，miR-20a在hASCs成骨分化中的作用仍不清楚。在本研究中，我们系统评估了miR-20a在体外调控hASCs成骨过程中的功能。用miR-20a过表达和miR-20a海绵慢病毒载体转导hASCs，并以绿色荧光蛋白（GFP）作为对照。结果显示，hASCs矿化后miR-20a转录上调。与miR-20a海绵组、GFP组和hASCs组相比，miR-20a过表达组在第7天和第14天显示出更高的碱性磷酸酶（ALP）活性。此外，在第14天，miR-20a过表达组中ALP的mRNA水平显著增加。此外，靶基因PPARγ的蛋白水平降低，而成骨分化相关蛋白ALP、骨钙素和RUNX2上调。附着于HA/β-TCP的hASCs呈现健康的多边形形态并形成细胞质延伸。miR-20a促进了细胞支架的成骨分化。综上所述，这些数据证实miRNA-20a在体外促进hASCs的成骨作用，其在体内的重要作用有待进一步研究。
