Cytoskeletal binding proteins distinguish cultured dental follicle cells and periodontal ligament cells
Cytoskeletal binding proteins distinguish cultured dental follicle cells and periodontal ligament cells
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细胞骨架结合蛋白区分培养的牙囊细胞和牙周膜细胞
DOI：
10.1016/j.yexcr.2015.12.011
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发表时间：
2016-07-01
影响因子：
3.7
通讯作者：
Tian, Weidong
中科院分区：
文献类型：
--
作者：
Li, Jie;Li, Hui;Tian, Weidong
人牙囊细胞（DFC）和牙周膜细胞（PDLC）来源于外胚间充质组织，具有干/祖细胞特性和诱导组织再生的能力。牙囊中的干细胞可分化为成牙骨质细胞、牙周膜成纤维细胞和成骨细胞，分别形成牙骨质、牙周膜和牙槽骨。虽然牙囊中的干细胞是牙周膜成纤维细胞的前体细胞，但区分培养的DFC和PDLCs的分子变化仍然未知。在本研究中，我们比较了两种细胞系的免疫表型特征和细胞周期状态。结果表明，DFCs和PDLCs在免疫表型和细胞周期方面表现出相似的特征。然后，我们采用同量异序标签相对和绝对定量（iTRAQ）蛋白质组学策略，以揭示两种细胞类型之间的分子差异。共鉴定了2138种蛋白质，其中39种蛋白质在DFC和PDLC之间始终差异表达。基因本体分析显示，PDLCs中高表达的蛋白亚群与肌动蛋白结合、细胞骨架蛋白结合和肌肉结构成分有关。经实时PCR、蛋白质印迹和免疫荧光染色验证。原肌球蛋白1（TPM 1）和钙调蛋白1（CALD 1）在PDLC中的表达高于DFC。我们的研究结果表明，PDLCs显示增强肌动蛋白细胞骨架动力学相对于DFC，而DFC可能表现出更强大的抗氧化防御能力相对于PDLCs。这项研究扩展了我们对培养的DFC和PDLCs蛋白质组的了解，并为在每种细胞类型中观察到的不同生物学特征的可能机制提供了新的见解。(C)2016由Elsevier Inc.出版
Human dental follicle cells (DFCs) and periodontal ligament cells (PDLCs) derived from the ectomesenchymal tissue, have been shown to exhibit stem/progenitor cell properties and the ability to induce tissue regeneration. Stem cells in dental follicle differentiate into cementoblasts, periodontal ligament fibroblasts and osteoblasts, these cells form cementum, periodontal ligament and alveolar bone, respectively. While stem cells in dental follicle are a precursor to periodontal ligament fibroblasts, the molecular changes that distinguish cultured DFCs from PDLCs are still unknown. In this study, we have compared the immunophenotypic features and cell cycle status of the two cell lines. The results suggest that DFCs and PDLCs displayed similar features related to immunophenotype and cell cycle. Then we employed an isobaric tag for relative and absolute quantitation (iTRAQ) proteomics strategy to reveal the molecular differences between the two cell types. A total of 2138 proteins were identified and 39 of these proteins were consistently differentially expressed between DFCs and PDLCs. Gene ontology analyses revealed that the protein subsets expressed higher in PDLCs were related to actin binding, cytoskeletal protein binding, and structural constituent of muscle. Upon validation by real-time PCR, western blotting, and immunofluorescence staining. Tropomyosin 1 (TPM1) and caldesmon 1 (CALD1) were expressed higher in PDLCs than in DFCs. Our results suggested that PDLCs display enhanced actin cytoskeletal dynamics relative to DFCs while DFCs may exhibit a more robust antioxidant defense ability relative to PDLCs. This study expands our knowledge of the cultured DFCs and PDLCs proteome and provides new insights into possible mechanisms responsible for the different biological features observed in each cell type. (C) 2016 Published by Elsevier Inc.