Liu Z H, Qi D D, Li X, Zhang S Q, Zhao Y, Fu L X, Lu L Y
Department of Spinal Surgery, East Hospital, Tongji University School of Medicine, Shanghai, China.
Department of Orthopedics, South of Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, China.
J Biol Regul Homeost Agents. 2021 Mar-Apr;35(2):605-615. doi: 10.23812/20-391-A.
Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is vital for bone formation, and its dysfunction is linked to osteoporosis (OP). In this work, we explored the function of long non-coding RNA (lncRNA) small nucleolar RNA host gene 14 (SNHG14) in regulating osteogenic differentiation of hBMSCs. In the present study, the expression of SNHG14 in hBMSCs obtained from OP patients was measured by quantitative real-time polymerase chain reaction (qRT-PCR). SNHG14 was over-expressed or knocked down in hBMSCs, and the expression levels of OP-related genes (ALP, OCN, and OPN) in hBMSCs were detected by qRT-PCR and Western blot. StarBase database and miRanda database were used to predict the binding sites between SNHG14 and miR-185-5p, and between miR-185-5p and 3'UTR of WNT1 inducible signaling pathway protein 2 (WISP2), respectively. Luciferase reporter gene assay was used to validate the binding relationship between SNHG14 and miR-185-5p, and miR-185-5p and 3'UTR of WISP2, respectively. Here, we report that SNHG14 was significantly down-regulated in hBMSCs obtained from patients with OP. Overexpression of SNHG14 promoted osteogenic differentiation, while knockdown of SNHG14 worked oppositely. Mechanistically, miR-185-5p was demonstrated to be a target of SNHG14, and could reverse the function of SNHG14. Additionally, WISP2 was identified as a target gene of miR-185-5p in hBMSCs and could be indirectly regulated by SNHG14. Taken together, down-regulation of SNHG14 in hBMSCs accelerated the progression of OP via regulating miR-185-5p/WISP2 axis.
人骨髓间充质干细胞（hBMSCs）的成骨分化对骨形成至关重要，其功能障碍与骨质疏松症（OP）相关。在本研究中，我们探究了长链非编码RNA（lncRNA）小核仁RNA宿主基因14（SNHG14）在调节hBMSCs成骨分化中的作用。在本研究中，通过定量实时聚合酶链反应（qRT-PCR）检测了OP患者来源的hBMSCs中SNHG14的表达。在hBMSCs中过表达或敲低SNHG14，通过qRT-PCR和蛋白质免疫印迹法检测hBMSCs中OP相关基因（碱性磷酸酶（ALP）、骨钙素（OCN）和骨桥蛋白（OPN））的表达水平。分别使用StarBase数据库和miRanda数据库预测SNHG14与miR-185-5p之间以及miR-185-5p与WNT1诱导信号通路蛋白2（WISP2）的3'非翻译区（3'UTR）之间的结合位点。分别使用荧光素酶报告基因实验验证SNHG14与miR-185-5p以及miR-185-5p与WISP2的3'UTR之间的结合关系。在此，我们报道OP患者来源的hBMSCs中SNHG14显著下调。SNHG14的过表达促进成骨分化，而敲低SNHG14则起相反作用。机制上，miR-185-5p被证明是SNHG14的靶标，并且可以逆转SNHG14的功能。此外，WISP2被鉴定为hBMSCs中miR-185-5p的靶基因，并且可以被SNHG14间接调节。综上所述，hBMSCs中SNHG14的下调通过调节miR-185-5p/WISP2轴加速了OP的进展。
