Department of Orthopedics, Yunyang County Peoples' Hospital, Chongqing 404500, China.
Department of Orthopaedics, Xi'an Ninth Hospital, No.151, East Section of South Second Ring Road, Beilin District, Xi'an, Shaanxi, 710054, China.
Crit Rev Eukaryot Gene Expr. 2023;33(3):61-70. doi: 10.1615/CritRevEukaryotGeneExpr.2022044929.
This study aimed to identify the possible function and the molecular mechanism of hsa_circ_0007334 in human bone marrow mesenchymal stem cells (hBMSCs) osteogenic differentiation. The level of hsa_circ_0007334 was detected by means of quantitative real-time polymerase chain reaction (RT-qPCR). Alkaline phosphatase (ALP), RUNX2, osterix (OSX), and osteocalcin (OCN) were monitored to analyze the degree of osteogenic differentiation under routine culture or under the control of hsa_circ_0007334. The proliferation of hBMSCs was tested with a cell counting kit-8 (CCK-8) assay. The migration of hBMSCs was tested using the Transwell assay. Bioinformatics analysis was used to predict the possible targets of hsa_circ_0007334 or miR-144-3p. Dual-luciferase reporter assay system was used to analyze the combination between hsa_circ_0007334 and miR-144-3p. Hsa_circ_0007334 was upregulated in osteogenic differentiation of hBMSCs. Osteogenic differentiation increased by hsa_circ_0007334 in vitro was confirmed with levels of ALP and bone markers (RUNX2, OCN, OSX). hsa_circ_0007334 overexpression promoted osteogenic differentiation, proliferation, and migration of hBMSCs, and knockdown of hsa_circ_0007334 has the opposite effects. miR-144-3p was identified as the target of hsa_circ_0007334. The targeting genes of miR-144-3p are involved in osteogenic-differentia-tion-related biological processes (such as bone development, epithelial cell proliferation, and mesenchymal cell apoptotic prosess) and pathways (including FoxO and VEGF signaling pathway). Hsa_circ_0007334, therefore, presents itself as a promising biological for osteogenic differentiation.
本研究旨在鉴定 hsa_circ_0007334 在人骨髓间充质干细胞（hBMSCs）成骨分化中的可能功能和分子机制。通过实时定量聚合酶链反应（RT-qPCR）检测 hsa_circ_0007334 的水平。在常规培养或 hsa_circ_0007334 控制下，监测碱性磷酸酶（ALP）、RUNX2、osterix（OSX）和骨钙素（OCN）的水平，以分析成骨分化程度。使用细胞计数试剂盒-8（CCK-8）测定 hBMSCs 的增殖。使用 Transwell 测定法检测 hBMSCs 的迁移。生物信息学分析用于预测 hsa_circ_0007334 或 miR-144-3p 的可能靶标。双荧光素酶报告基因检测系统用于分析 hsa_circ_0007334 和 miR-144-3p 之间的结合。hsa_circ_0007334 在 hBMSCs 的成骨分化中上调。通过 hsa_circ_0007334 在体外促进成骨分化，ALP 和骨标志物（RUNX2、OCN、OSX）水平得到证实。hsa_circ_0007334 的过表达促进了 hBMSCs 的成骨分化、增殖和迁移，而 hsa_circ_0007334 的敲低则产生相反的效果。miR-144-3p 被鉴定为 hsa_circ_0007334 的靶标。miR-144-3p 的靶基因参与成骨分化相关的生物过程（如骨发育、上皮细胞增殖和间充质细胞凋亡过程）和途径（包括 FoxO 和 VEGF 信号通路）。因此，hsa_circ_0007334 作为一种有前途的成骨分化生物。
