Department of Prosthodontics, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
Department of Anatomy, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand.
Oral Dis. 2019 May;25(4):1203-1213. doi: 10.1111/odi.13065. Epub 2019 Mar 5.
Jagged1 regulates several biological functions in human periodontal ligament cells (hPDLs). The present study aimed to evaluate mRNA expression profiling of Jagged1-treated hPDLs using microarray technique.
Notch ligands, Jagged1, were indirectly immobilized on tissue culture surface. Subsequently, hPDLs were seeded on Jagged1 immobilized surface and maintained in growth medium for 48 hr. Total RNA was collected and processed. Gene expression profiling was examined using microarray technique. Real-time polymerase chain reaction and immunofluorescence staining were employed to determine mRNA and protein expression levels, respectively. Cell proliferation and colony-forming unit assay were performed. Cell cycle was evaluated using propidium iodide staining and flow cytometry analysis.
The isolated cells demonstrated fibroblast-like morphology and exhibited the co-expression of CD44, CD90, and CD105 surface markers. After stimulated with Jagged1, the total of 411 genes was differentially expressed, consisting both coding and non-coding genes. For coding genes, 165 and 160 coding genes were upregulated and downregulated, respectively. Pathway analysis revealed that the upregulated genes were mainly involved in cellular interactions, signal transduction, and collagen formation and degradation while the downregulated genes were in the events and phases in cell cycle. Jagged1 significantly decreased cell proliferation, reduced colony-forming unit ability, and induced G0/G1 cell cycle arrest in hPDLs.
Jagged1 regulates various biological pathways in hPDLs. This gene expression profiling could help to understand the mechanisms potentially involved in the Notch signaling regulation in periodontal homeostasis.
Jagged1 调控人牙周膜细胞（hPDLs）的多种生物学功能。本研究旨在通过微阵列技术评估 Jagged1 处理的 hPDLs 的 mRNA 表达谱。
将 Notch 配体 Jagged1 间接固定在组织培养表面上。随后，将 hPDLs 接种到 Jagged1 固定化表面上，并在生长培养基中维持 48 小时。收集总 RNA 并进行处理。使用微阵列技术检查基因表达谱。实时聚合酶链反应和免疫荧光染色分别用于确定 mRNA 和蛋白质表达水平。进行细胞增殖和集落形成单位测定。通过碘化丙啶染色和流式细胞术分析评估细胞周期。
分离的细胞呈现出成纤维细胞样形态，并表现出 CD44、CD90 和 CD105 表面标志物的共表达。用 Jagged1 刺激后，总共有 411 个基因差异表达，包括编码和非编码基因。对于编码基因，分别有 165 个和 160 个编码基因上调和下调。通路分析显示，上调基因主要参与细胞相互作用、信号转导以及胶原形成和降解，而下调基因则与细胞周期中的事件和阶段有关。Jagged1 显著降低 hPDLs 的细胞增殖、减少集落形成单位能力，并诱导 G0/G1 细胞周期停滞。
Jagged1 调控 hPDLs 中的多种生物学途径。该基因表达谱有助于理解 Notch 信号转导在牙周稳态调节中可能涉及的机制。
