转胶蛋白通过介导转化生长因子-β1 促进人牙周膜细胞的增殖。
Transgelin mediates transforming growth factor-β1-induced proliferation of human periodontal ligament cells.
机构信息
Department of Endodontology and Operative Dentistry, Faculty of Dental Science, Kyushu University, Fukuoka, Japan.
Division of General Dentistry, Kyushu University Hospital, Kyushu University, Fukuoka, Japan.
出版信息
J Periodontal Res. 2017 Dec;52(6):984-993. doi: 10.1111/jre.12466. Epub 2017 Jun 7.
BACKGROUND AND OBJECTIVE
Human periodontal ligament cells (HPDLCs) express transforming growth factor-β1 (TGF-β1) that regulates differentiation and proliferation, and plays key roles in homeostasis of PDL tissue. Transgelin is a cytoskeleton-associated protein with an Smad-binding element in its gene promoter region. In this study, we examined the localization and potential function of transgelin in PDL tissue and cells.
MATERIAL AND METHODS
Microarray analysis of HPDLC lines (2-14, 2-23 and 2-52) was performed. Expression of transgelin in HPDLCs was examined by quantitative reverse transcription-polymerase chain reaction, immunofluorescence staining and western blot analysis. Effects of TGF-β1 and its signaling inhibitor, SB431542, on transgelin expression in HPDLCs were examined by western blot analysis. The effects of transgelin knockdown by small interfering RNA (siRNA) on HPDLC proliferation stimulated by TGF-β1 were assessed by WST-1 assay.
RESULTS
In microarray and quantitative reverse transcription-polymerase chain reaction analyses, the expression levels of transgelin (TAGLN) in 2-14 and 2-23 cells, which highly expressed PDL markers such as periostin (POSTN), tissue non-specific alkaline phosphatase (ALPL), α-smooth muscle actin (ACTA2) and type I collagen A1 (COL1A1), was significantly higher than those in 2-52 cells that expressed PDL markers weakly. Immunohistochemical and immunofluorescence staining revealed expression of transgelin in rat PDL tissue and HPDLCs. In HPDLCs, TGF-β1 treatment upregulated transgelin expression, whereas inhibition of the type 1 TGF-β1 receptor by SB431542 suppressed this upregulation. Furthermore, TAGLN siRNA transfection did not promote the proliferation of HPDLCs treated with TGF-β1. The expression levels of CCNA2 and CCNE1, which regulate DNA synthesis and mitosis through the cell cycle, were also not upregulated in HPDLCs transfected with TAGLN siRNA.
CONCLUSION
Transgelin is expressed in PDL tissue and might have a role in HPDLC proliferation induced by TGF-β1 stimulation.
背景与目的
人牙周韧带细胞（HPDLCs）表达转化生长因子-β1（TGF-β1），该因子调节分化和增殖，在牙周组织的稳态中发挥关键作用。转凝胶蛋白是一种细胞骨架相关蛋白，其基因启动子区域含有 Smad 结合元件。本研究旨在检测转凝胶蛋白在牙周组织和细胞中的定位和潜在功能。
材料与方法
对 HPDLC 系（2-14、2-23 和 2-52）进行微阵列分析。通过定量逆转录聚合酶链反应、免疫荧光染色和 Western blot 分析检测 HPDLC 中转凝胶蛋白的表达。通过 Western blot 分析检测 TGF-β1及其信号抑制剂 SB431542 对 HPDLC 中转凝胶蛋白表达的影响。通过 WST-1 测定评估转凝胶蛋白 siRNA 敲低对 TGF-β1 刺激的 HPDLC 增殖的影响。
结果
在微阵列和定量逆转录聚合酶链反应分析中，在 2-14 和 2-23 细胞中转凝胶蛋白（TAGLN）的表达水平明显高于在 2-52 细胞中的表达水平，2-14 和 2-23 细胞高度表达牙周标志物，如骨桥蛋白（POSTN）、组织非特异性碱性磷酸酶（ALPL）、α-平滑肌肌动蛋白（ACTA2）和 I 型胶原 A1（COL1A1）。免疫组织化学和免疫荧光染色显示转凝胶蛋白在大鼠牙周组织和 HPDLC 中的表达。在 HPDLC 中，TGF-β1 处理上调转凝胶蛋白的表达，而 SB431542 抑制 I 型 TGF-β1 受体抑制这种上调。此外，TAGLN siRNA 转染不会促进 TGF-β1 处理的 HPDLC 增殖。在转染 TAGLN siRNA 的 HPDLC 中，调节 DNA 合成和有丝分裂的细胞周期相关蛋白 CCNA2 和 CCNE1 的表达水平也没有上调。
结论
转凝胶蛋白在牙周组织中表达，可能在 TGF-β1 刺激诱导的 HPDLC 增殖中发挥作用。
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