Department of Orthopedics, Shanghai Eighth People's Hospital, Shanghai, 200235, China.
Department of Rehabilitation Medicine, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, 533000, Guangxi, China.
J Orthop Surg Res. 2024 Sep 16;19(1):572. doi: 10.1186/s13018-024-05053-8.
Osteoporosis results from decreased bone mass and disturbed bone structure. Human bone marrow mesenchymal stem cells (hBMSCs) demonstrate robust osteogenic differentiation, a critical process for bone formation. This research was designed to examine the functions of LINC01133 in osteogenic differentiation.
Differentially expressed lncRNAs affecting osteogenic differentiation in hBMSCs were identified from the GEO database. A total of 74 osteoporosis patients and 70 controls were enrolled. hBMSCs were stimulated to undergo osteogenic differentiation using an osteogenic differentiation medium (OM). RT-qPCR was performed to evaluate LINC01133 levels and osteogenesis-related genes such as osteocalcin, osteopontin, and RUNX2. An alkaline phosphates (ALP) activity assay was conducted to assess osteogenic differentiation. Cell apoptosis was detected using flow cytometry. Dual luciferase reporter assay and RIP assay were employed to investigate the association between miR-214-3p and LINC01133 or CTNNB1. Loss or gain of function assays were conducted to elucidate the impact of LINC01133 and miR-214-3p on osteogenic differentiation of hBMSCs.
LINC01133 and CTNNB1 expression decreased in osteoporotic patients but increased in OM-cultured hBMSCs, whereas miR-214-3p showed an opposite trend. Depletion of LINC01133 suppressed the expression of genes associated with bone formation and ALP activity triggered by OM in hBMSCs, leading to increased cell apoptosis. Nevertheless, this suppression was partially counteracted by the reduced miR-214-3p levels. Mechanistically, LINC01133 and CTNNB1 were identified as direct targets of miR-214-3p.
Our study highlights the role of LINC01133 in positively regulating CTNNB1 expression by inhibiting miR-214-3p, thereby promoting osteogenic differentiation of BMSCs. These findings may provide valuable insights into bone regeneration in osteoporosis.
骨质疏松症是由骨量减少和骨结构紊乱引起的。人骨髓间充质干细胞（hBMSCs）具有强大的成骨分化能力，这是骨形成的关键过程。本研究旨在探讨 LINC01133 在成骨分化中的作用。
从 GEO 数据库中鉴定出影响 hBMSCs 成骨分化的差异表达 lncRNA。共纳入 74 例骨质疏松症患者和 70 例对照。使用成骨分化培养基（OM）刺激 hBMSCs 进行成骨分化。采用 RT-qPCR 检测 LINC01133 水平及骨钙素、骨桥蛋白和 RUNX2 等成骨相关基因。碱性磷酸酶（ALP）活性测定评估成骨分化。采用流式细胞术检测细胞凋亡。双荧光素酶报告基因和 RIP 测定用于研究 miR-214-3p 与 LINC01133 或 CTNNB1 之间的关联。通过缺失或获得功能实验阐明 LINC01133 和 miR-214-3p 对 hBMSCs 成骨分化的影响。
骨质疏松症患者中 LINC01133 和 CTNNB1 的表达降低，而 OM 培养的 hBMSCs 中表达升高，而 miR-214-3p 则呈现相反的趋势。LINC01133 耗竭抑制了 OM 诱导的 hBMSCs 中成骨相关基因的表达和 ALP 活性，导致细胞凋亡增加。然而，这种抑制作用部分被降低的 miR-214-3p 水平所抵消。机制上，LINC01133 和 CTNNB1 被鉴定为 miR-214-3p 的直接靶点。
本研究强调了 LINC01133 通过抑制 miR-214-3p 正向调控 CTNNB1 表达，从而促进 BMSCs 成骨分化的作用。这些发现可能为骨质疏松症中的骨再生提供有价值的见解。
