Department of Joint Surgery, Yantai Yuhuangding Hospital, Yantai, Shandong 264000, P.R. China.
Department of Orthopedics, Pingdu People's Hospital, Pingdu, Shandong 266700, P.R. China.
Mol Med Rep. 2020 Jan;21(1):43-50. doi: 10.3892/mmr.2019.10791. Epub 2019 Nov 5.
Mesenchymal stem cells (MSCs) are used to investigate regeneration and differentiation. MicroRNA‑204 (miR‑204) in involved in the Runt‑related transcription factor 2/alkaline phosphatase/bone morphogenic protein 2 (Runx2/ALP/BMP2) signaling pathway that regulates bone marrow mesenchymal stem cell (BMSC) differentiation; however, the mechanisms underlying the effects of miR‑204 are yet to be determined. The aim of the present study was to investigate the effects of miR‑204 on BMSC differentiation. BMSCs were derived from rat bone marrow. The expression levels of Runx2, ALP and BMP2 were measured via reverse transcription‑quantitative polymerase chain reaction and western blot analyses following transfection of BMSCs with miR‑204 agomir or BMP2 expression vector. The ability of the miR‑204 gene to directly bind BMP2 mRNA was assessed using dual‑luciferase assays. Ossification was measured via alizarin red stain assays. It was observed that the expression levels of Runx2 and ALP increased over time, whereas those of miR‑204 decreased; additionally, miR‑204 agomir upregulation inhibited the expression of Runx2, ALP and BMP2 in BMSCs. It was revealed that miR‑204 directly interacted with BMP2 mRNA, and that transfection with miR‑204 agomir suppressed ossification in BMSCs by targeting the BMP2/Runx2/ALP signaling pathway.
间充质干细胞(MSCs)用于研究再生和分化。MicroRNA-204(miR-204)参与 Runt 相关转录因子 2/碱性磷酸酶/骨形态发生蛋白 2(Runx2/ALP/BMP2)信号通路,该通路调节骨髓间充质干细胞(BMSC)分化;然而,miR-204 作用的机制尚待确定。本研究旨在探讨 miR-204 对 BMSC 分化的影响。BMSCs 来源于大鼠骨髓。转染 BMSCs 后,通过逆转录-定量聚合酶链反应和 Western blot 分析测量 Runx2、ALP 和 BMP2 的表达水平miR-204 agomir 或 BMP2 表达载体。使用双荧光素酶测定评估 miR-204 基因与 BMP2 mRNA 直接结合的能力。通过茜素红染色测定骨化。结果观察到 Runx2 和 ALP 的表达水平随时间增加而增加,而 miR-204 的表达水平降低;此外,miR-204 agomir 的上调抑制了 BMSCs 中 Runx2、ALP 和 BMP2 的表达。结果表明,miR-204 与 BMP2 mRNA 直接相互作用,转染 miR-204 agomir 通过靶向 BMP2/Runx2/ALP 信号通路抑制 BMSCs 的骨化。