Wang Qiong, Wang Junfang, Gu Xiaofeng, Feng Dehong, Li Ding, Jiang Tao
Department of Clinical Laboratory, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
Department of Orthopedics, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi, Jiangsu 214000, P.R. China.
Exp Ther Med. 2021 Jul;22(1):725. doi: 10.3892/etm.2021.10157. Epub 2021 May 4.
MicroRNA (miRNA/miR)-124-3p has been extensively studied in tumor biology and stem cells. However, little is known regarding its functional roles in the differentiation of precartilaginous stem cells (PSCs) into nucleus pulposus-like cells (NPLCs). In the present study, using miRNA microarray screening, it was demonstrated that the miRNA expression profiles differed between rat primary PSCs and TGF-β1-induced differentiated NPLCs, and that miR-124-3p was significantly differentially expressed during the differentiation of PSCs to NPLCs. Furthermore, RT-qPCR analysis verified that miR-124-3p expression was decreased during PSC differentiation, with the lowest levels being detected at the later stages. Subsequent experiments revealed that miR-124-3p overexpression significantly decreased the expression of the extracellular matrix proteins, aggrecan and collagen type II, which was accompanied by a significant decrease in follistatin-related protein 1 (FSTL1) expression levels. Moreover, bioinformatics analysis indicated that FSTL1 was a potential target of miR-124-3p, which was additionally verified using luciferase reporter assays. Taken together, these data revealed a specific regulatory pathway of miR-124-3p, which negatively regulated its target gene, FSTL1, during the differentiation of PSCs to NPLCs, and suggested a functional role for miR-124-3p in the differentiation of PSCs.
微小RNA（miRNA/miR）-124-3p已在肿瘤生物学和干细胞领域得到广泛研究。然而，关于其在软骨前体细胞（PSC）向髓核样细胞（NPLC）分化过程中的功能作用，人们所知甚少。在本研究中，通过miRNA微阵列筛选发现，大鼠原代PSC与转化生长因子-β1诱导分化的NPLC之间的miRNA表达谱存在差异，且miR-124-3p在PSC向NPLC分化过程中显著差异表达。此外，逆转录定量聚合酶链反应（RT-qPCR）分析证实，miR-124-3p在PSC分化过程中表达降低，在后期检测到的水平最低。随后的实验表明，miR-124-3p过表达显著降低了细胞外基质蛋白聚集蛋白聚糖和II型胶原蛋白的表达，同时卵泡抑素相关蛋白1（FSTL1）的表达水平也显著降低。此外，生物信息学分析表明FSTL1是miR-124-3p的潜在靶标，荧光素酶报告基因实验进一步验证了这一点。综上所述，这些数据揭示了miR-124-3p的特定调控途径，即在PSC向NPLC分化过程中负调控其靶基因FSTL1，并提示miR-124-3p在PSC分化中具有功能作用。
