Department of Orthopedic Surgery, Second Affiliated Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia Autonomous Region 010030, P.R. China.
Department of Orthopedic Surgery, Inner Mongolia Hospital of Traditional Chinese Medicine, Hohhot, Inner Mongolia Autonomous Region 010010, P.R. China.
Mol Med Rep. 2022 Jul;26(1). doi: 10.3892/mmr.2022.12750. Epub 2022 May 26.
Osteonecrosis of the femoral head (ONFH) affects the life of patients. MicroRNA‑141 (miR‑141) has been found associated with proliferation of bone marrow‑derived mesenchymal stem cells (BMSCs). E2F transcription factor 3 (E2F3) has been identified as the target of miR‑141 to regulate cell proliferation. The aim of the present study was to investigate whether miR‑141 and E2F3 were involved in the osteogenic differentiation of BMSCs during ONFH. BMSCs from 4‑week‑old Sprague‑Dawley rats were transduced with miR‑141 mimic or inhibitor lentiviruses. Alkaline phosphatase staining was performed to confirm osteogenic differentiation. Reverse transcription‑quantitative PCR, luciferase reporter assays and western blot analysis were also used to examine the interaction between E2F3 and miR‑141 in BMSCs from the control and ONFH rats. The lentiviral transductions were carried out successfully. The mRNA expression levels of miR‑141 in ONFH were upregulated, while those of E2F3 were downregulated compared with the control rat. The luciferase reporter assays indicated that miR‑141 could target E2F3. miR‑141 knockdown upregulated the mRNA expression levels of E2F3. In addition, osteogenic differentiation of BMSCs was inhibited following miR‑141 overexpression, but increased following miR‑141 knockdown, as evidenced by the results of the alkaline phosphatase staining and western blot analysis. In conclusion, miR‑141 inhibits the osteogenic differentiation of BMSCs in ONFH by targeting E2F3. These two molecules may represent novel candidates to examine in order to investigate the mechanism underlying ONFH.
股骨头坏死(ONFH)影响患者的生活。已经发现 microRNA-141(miR-141)与骨髓间充质干细胞(BMSCs)的增殖有关。E2F 转录因子 3(E2F3)已被确定为 miR-141 的靶标,以调节细胞增殖。本研究旨在探讨 miR-141 和 E2F3 是否参与 ONFH 期间 BMSCs 的成骨分化。用 miR-141 模拟物或抑制剂慢病毒转导 4 周龄 Sprague-Dawley 大鼠的 BMSCs。进行碱性磷酸酶染色以确认成骨分化。还使用逆转录-定量 PCR、荧光素酶报告基因测定和 Western blot 分析来检查对照和 ONFH 大鼠 BMSCs 中 E2F3 和 miR-141 之间的相互作用。慢病毒转导成功进行。与对照组大鼠相比,ONFH 大鼠中 miR-141 的 mRNA 表达水平上调,而 E2F3 的 mRNA 表达水平下调。荧光素酶报告基因测定表明 miR-141 可以靶向 E2F3。miR-141 下调后,E2F3 的 mRNA 表达水平上调。此外,miR-141 过表达抑制 BMSCs 的成骨分化,但 miR-141 下调后增加,碱性磷酸酶染色和 Western blot 分析的结果证实了这一点。总之,miR-141 通过靶向 E2F3 抑制 ONFH 中 BMSCs 的成骨分化。这两个分子可能代表研究 ONFH 机制的新候选物。