Shanghai General Hospital of Nanjing Medical University, Shanghai 200080, P.R. China.
Department of Orthopedics, Shanghai Jiahui International Hospital, Shanghai 200233, P.R. China.
Int J Mol Med. 2020 Aug;46(2):709-717. doi: 10.3892/ijmm.2020.4615. Epub 2020 May 22.
Accumulating evidence has documented that microRNAs (miRNAs or miRs) function as important post‑transcriptional regulators of the differentiation of mesenchymal stem cells (MSCs), including human adipose‑derived mesenchymal stem cells (hADSCs); however, their roles in hADSC osteogenic differentiation require further investigation. The present study aimed to investigate the role of miRNAs in the osteogenic differentiation of hADSCs and to elucidate the underlying molecular mechanisms. Using an miRNA microarray, it was found that 24 miRNAs were upregulated and 14 miRNAs were downregulated compared with the undifferentiated cells, and miR‑29b‑3p (miR‑29b) was selected for further experiments. Functional experiments revealed that the upregulation of miR‑29b by agomir‑29b significantly enhanced alkaline phosphatase (ALP) activity and the mineralization of extracellular matrix (ECM), and led to an increase in the mRNA and protein levels of osteogenic marker genes, including runt‑related transcription factor 2 (Runx2), osteopontin (OPN), osteocalcin (OCN) and bone sialoprotein (BSP), whereas the knockdown of miR‑29b suppressed these processes. In addition, phosphatase and tensin homolog (PTEN), a negative regulator of the AKT/β‑catenin pathway, was identified as a direct target of miR‑29b in the hADSCs. Moreover, it was observed that the overexpression of miR‑29b activated the AKT/β‑catenin signaling pathway by inhibiting PTEN expression in the hADSCs. Most importantly, it was also found that the overexpression of PTEN reversed the promoting effects of miR‑29b on osteogenic differentiation. On the whole, these findings suggest that miR‑29b promotes the osteogenic differentiation of hADSCs by modulating the PTEN/AKT/β‑catenin signaling pathway. Thus, this miRNA may be a promising target for the active modulation of hADSC‑derived osteogenesis.
越来越多的证据表明,微小 RNA(miRNA 或 miR)作为重要的转录后调节剂,可调节间充质干细胞(MSCs)的分化,包括人脂肪来源间充质干细胞(hADSCs);然而,其在 hADSC 成骨分化中的作用仍需进一步研究。本研究旨在探讨 miRNA 在 hADSC 成骨分化中的作用,并阐明其潜在的分子机制。通过 miRNA 微阵列分析,发现与未分化细胞相比,有 24 个 miRNA 上调,14 个 miRNA 下调,选择 miR-29b-3p(miR-29b)进行进一步实验。功能实验表明,agomir-29b 上调 miR-29b 可显著增强碱性磷酸酶(ALP)活性和细胞外基质(ECM)的矿化,并导致成骨标志物基因,包括 runt 相关转录因子 2(Runx2)、骨桥蛋白(OPN)、骨钙素(OCN)和骨涎蛋白(BSP)的 mRNA 和蛋白水平增加,而 miR-29b 的敲低则抑制了这些过程。此外,发现磷酸酶和张力蛋白同源物(PTEN)是 AKT/β-连环蛋白通路的负调节剂,是 hADSCs 中 miR-29b 的直接靶标。此外,观察到 miR-29b 通过抑制 hADSCs 中 PTEN 的表达,激活 AKT/β-连环蛋白信号通路。最重要的是,还发现过表达 PTEN 可逆转 miR-29b 对成骨分化的促进作用。综上所述,这些发现表明,miR-29b 通过调节 PTEN/AKT/β-连环蛋白信号通路促进 hADSC 的成骨分化。因此,这种 miRNA 可能是调节 hADSC 源性成骨的有前途的靶点。