Liang Daning, Song Guodong, Zhang Zhenning
Department of Medical Beauty, Shenzhen Hospital (Guangming), University of Chinese Academy of Sciences, Shenzhen, Guangdong 518107, P.R. China.
Department of Craniomaxillofacial Surgery, Plastic Surgery Hospital of Chinese Academy of Medical Sciences, Beijing 100144, P.R. China.
Exp Ther Med. 2022 Apr;23(4):309. doi: 10.3892/etm.2022.11238. Epub 2022 Feb 24.
The current study aimed to investigate the potential function and mechanism of microRNA (miR)-216a-3p in the osteogenic differentiation of human adipose-derived stem cells (hADSCs). Dynamic expression changes of miR-216a-3p in the osteogenic differentiation of hADSCs were examined by reverse transcription-quantitative PCR (RT-qPCR). Regulatory effects of miR-216a-3p on the relative levels of osteogenesis-associated genes were also detected by RT-qPCR and western blotting. The relationship between miR-216a-3p and Wnt3a was verified through a dual-luciferase reporter assay. Furthermore, the influence of miR-216a-3p on the Wnt/β-catenin signaling pathway during the osteogenic differentiation of hADSCs was investigated by western blotting. The results revealed that during the osteogenic differentiation process of hADSCs, miR-216a-3p was downregulated and Wnt3a was upregulated. It was further verified that Wnt3a was the target of miR-216a-3p. Through inactivation of the Wnt/β-catenin signaling pathway, miR-216a-3p was able to mediate osteogenic differentiation of hADSCs. In conclusion, by targeting Wnt3a, miR-216a-3p mediated the osteogenic differentiation of hADSCs, which negatively regulated the Wnt/β-catenin signaling pathway.
本研究旨在探讨微小RNA(miR)-216a-3p在人脂肪来源干细胞(hADSCs)成骨分化中的潜在功能及机制。通过逆转录定量聚合酶链反应(RT-qPCR)检测miR-216a-3p在hADSCs成骨分化过程中的动态表达变化。还通过RT-qPCR和蛋白质免疫印迹法检测miR-216a-3p对成骨相关基因相对水平的调控作用。通过双荧光素酶报告基因检测验证miR-216a-3p与Wnt3a之间的关系。此外,通过蛋白质免疫印迹法研究miR-216a-3p对hADSCs成骨分化过程中Wnt/β-连环蛋白信号通路的影响。结果显示,在hADSCs成骨分化过程中,miR-216a-3p表达下调,Wnt3a表达上调。进一步验证Wnt3a是miR-216a-3p的靶标。通过使Wnt/β-连环蛋白信号通路失活,miR-216a-3p能够介导hADSCs的成骨分化。总之,miR-216a-3p通过靶向Wnt3a介导hADSCs的成骨分化,对Wnt/β-连环蛋白信号通路起负向调控作用。