Department of Orthopaedics, Affiliated Zhongshan Hospital of Dalian University, Dalian, 116001, Liaoning, People's Republic of China.
Department of Biomedical Engineering, Dalian University of Technology, Dalian, 116024, Liaoning, People's Republic of China.
Stem Cell Res Ther. 2021 Jan 7;12(1):18. doi: 10.1186/s13287-020-02075-x.
Steroid-induced osteonecrosis of the femoral head (SONFH) is a devastating orthopedic disease, which seriously affects the quality of life of patients. The study aims to investigate the effects of LncRNA NORAD on SONFH.
Human bone marrow-derived mesenchymal stem cells (hBMSCs) were isolated from the proximal femur of patients during routine orthopedic surgery and then cultured with dexamethasone (Dex) and transfected with NORAD overexpression vector, siRNA-NORAD and miR-26a-5p mimics. The mRNA expression of NORAD, miR-26a-5p, OPG, RANK, and RANKL was detected by RT-qPCR. Cell proliferation and apoptosis was measured by CCK-8 assay and flow cytometry, respectively. The protein expression of RUNX2, OPG, RANK, and RANKL was detected by western blot. The dual-luciferase reporter gene assay was performed to confirm the binding between NORAD and miR-26a-5p.
NORAD expression was downregulated in SONFH tissues, while miR-26a-5p expression was upregulated. Overexpression of NORAD improved DEX-induced inhibition of proliferation and differentiation, and promotion of apoptosis in hBMSCs, while knockdown of NORAD led to the opposite results. Moreover, NORAD improved DEX-induced inhibition of proliferation and differentiation, and promotion of apoptosis by regulation of miR-26a-5p in hBMSCs.
NORAD expression was downregulated in SONFH tissues, while miR-26a-5p expression was upregulated. NORAD improved DEX-induced inhibition of proliferation and differentiation, and promotion of apoptosis by regulation of miR-26a-5p in hBMSCs.
激素诱导性股骨头坏死（SONFH）是一种破坏性的骨科疾病，严重影响患者的生活质量。本研究旨在探讨 LncRNA NORAD 对 SONFH 的影响。
从常规骨科手术中患者的近端股骨分离人骨髓间充质干细胞（hBMSCs），并用地塞米松（Dex）培养，并转染 NORAD 过表达载体、siRNA-NORAD 和 miR-26a-5p 模拟物。通过 RT-qPCR 检测 NORAD、miR-26a-5p、OPG、RANK 和 RANKL 的 mRNA 表达。通过 CCK-8 测定和流式细胞术分别测量细胞增殖和凋亡。通过 Western blot 检测 RUNX2、OPG、RANK 和 RANKL 的蛋白表达。通过双荧光素酶报告基因实验证实 NORAD 与 miR-26a-5p 的结合。
SONFH 组织中 NORAD 表达下调，而 miR-26a-5p 表达上调。NORAD 的过表达改善了 DEX 诱导的 hBMSCs 增殖和分化抑制及凋亡促进，而 NORAD 的敲低则导致相反的结果。此外，NORAD 通过调节 hBMSCs 中的 miR-26a-5p 改善了 DEX 诱导的增殖和分化抑制及凋亡促进。
SONFH 组织中 NORAD 表达下调，而 miR-26a-5p 表达上调。NORAD 通过调节 miR-26a-5p 改善了 DEX 诱导的 hBMSCs 增殖和分化抑制及凋亡促进。
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