Wang Chengze, Dong Lingling, Wang Ying, Jiang Zhiwei, Zhang Jing, Yang Guoli
Key Laboratory of Oral Biomedical Research of Zhejiang Province, The Affiliated Hospital of Stomatology, School of Stomatology, Zhejiang University School of Medicine, Hangzhou, China.
Front Genet. 2021 Sep 27;12:750827. doi: 10.3389/fgene.2021.750827. eCollection 2021.
Human periodontal ligament cells (PDLCs) play an important role in periodontal tissue stabilization and function. In the process of osteogenic differentiation of PDLSCs, the regulation of molecular signal pathways are complicated. In this study, the sequencing results of three datasets on GEO were used to comprehensively analyze the miRNA-mRNA network during the osteogenic differentiation of PDLSCs. Using the GSE99958 and GSE159507, a total of 114 common differentially expressed genes (DEGs) were identified, including 62 up-regulated genes and 52 down-regulated genes. GO enrichment analysis was performed. The up-regulated 10 hub genes and down-regulated 10 hub genes were screened out by protein-protein interaction network (PPI) analysis and STRING in Cytoscape. Similarly, differentially expressed miRNAs (DEMs) were selected by limma package from GSE159508. Then, using the miRwalk website, we further selected 11 miRNAs from 16 DEMs that may have a negative regulatory relationship with hub genes. RT-PCR verification revealed that nine DEMs and 18 hub genes showed the same trend as the RNA-seq results during the osteogenic differentiation of PDLSCs. Finally, using miR-584-5p inhibitor and mimics, it was found that miR-584-5p negatively regulates the osteogenic differentiation of PDLSCs . In summary, the present results found several potential osteogenic-related genes and identified candidate miRNA-mRNA networks for the further study of osteogenic differentiation of PDLSCs.
人牙周膜细胞（PDLCs）在牙周组织的稳定和功能中发挥着重要作用。在牙周膜干细胞（PDLSCs）成骨分化过程中，分子信号通路的调控较为复杂。在本研究中，利用GEO上三个数据集的测序结果，全面分析了PDLSCs成骨分化过程中的miRNA-mRNA网络。利用GSE99958和GSE159507，共鉴定出114个共同差异表达基因（DEGs），其中上调基因62个，下调基因52个。进行了基因本体（GO）富集分析。通过Cytoscape中的蛋白质-蛋白质相互作用网络（PPI）分析和STRING筛选出上调的10个核心基因和下调的10个核心基因。同样，利用limma软件包从GSE159508中筛选出差异表达的miRNA（DEMs）。然后，通过miRwalk网站，我们从16个DEMs中进一步筛选出11个可能与核心基因存在负调控关系的miRNA。逆转录-聚合酶链反应（RT-PCR）验证显示，在PDLSCs成骨分化过程中，9个DEMs和18个核心基因的表达趋势与RNA测序结果一致。最后，使用miR-584-5p抑制剂和模拟物，发现miR-584-5p对PDLSCs的成骨分化具有负调控作用。综上所述，本研究结果发现了几个潜在的成骨相关基因，并确定了候选的miRNA-mRNA网络，为进一步研究PDLSCs的成骨分化提供了依据。
