Effects of Homeobox Gene Distal-less 3 on Proliferation and Odontoblastic Differentiation of Human Dental Pulp Cells
Effects of Homeobox Gene Distal-less 3 on Proliferation and Odontoblastic Differentiation of Human Dental Pulp Cells
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同源框基因distal-less 3对人牙髓细胞增殖和成牙本质细胞分化的影响
DOI：
10.1016/j.joen.2012.07.009
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发表时间：
2012-11-01
影响因子：
4.2
通讯作者：
Fan, Mingwen
中科院分区：
文献类型：
--
作者：
Li, Xiaoyan;Yang, Guobin;Fan, Mingwen
同源结构域基因Distal-less-3 （Dlx3）在牙齿发育过程中起着至关重要的作用。本研究旨在探讨Dlx3对人牙髓细胞（hDPCs）增殖和成牙细胞分化的影响。方法：用重组慢病毒感染人DPCs，稳定过表达Dlx3，研究Dlx3对hDPCs的生物学效应。采用直接细胞计数法和5-乙基-2′-脱氧尿苷掺入法检测hDPCs的增殖情况。采用von Kossa染色和碱性磷酸酶活性测定评价hDPCs的成牙分化。实时聚合酶链反应检测牙本质唾液磷蛋白（DSPP）、牙本质基质酸性磷蛋白1 （DMP1）、碱性磷酸酶（ALP）、巢蛋白（Nes）等重要矿物质基因。Western blot检测外源Dlx3存在或不存在时牙本质唾液磷蛋白（DSP）和DMP1蛋白表达的差异。结果：Dlx3过表达降低了hDPCs的增殖能力。Dlx3通过促进矿化结节形成来增强hDPCs的分化，上调ALP活性以及矿化相关基因DSPP、DMP1、ALP、Nes的表达。同时，在外源Dlx3存在下，DSP和DMP1蛋白水平显著升高。结论：Dlx3是hDPCs增殖和成牙细胞分化的有效调节剂。[J] end2012;38:1504-1510。
Introduction: Homeodomain gene Distal-less-3 (Dlx3) plays an essential role in tooth development. The aim of this study was to investigate the effects of Dlx3 on proliferation and odontoblastic differentiation of human dental pulp cells (hDPCs). Methods: Human DPCs were infected by recombinant lentivirus to overexpress Dlx3 stably, and the biological effects of Dlx3 on the hDPCs were investigated. Proliferation of the hDPCs was measured by direct cell counting and 5-ethynyl-2'-deoxyuridine incorporation assay. Odontogenic differentiation of hDPCs was evaluated by von Kossa staining and alkaline phosphatase activity assay. Important mineral genes such as dentin sialophosphoprotein (DSPP), dentin matrix acidic phosphoprotein 1 (DMP1), alkaline phosphatase (ALP), and nestin (Nes) were determined by real-time polymerase chain reaction. Western blot analysis was performed to determine the difference of expressions of protein of dentin sialophosphoprotein (DSP) and DMP1 with or without the presence of exogenous Dlx3. Results: Overexpression of Dlx3 decreased the proliferation ability of hDPCs. Dlx3 enhanced differentiation of hDPCs with promoting mineralization nodule formation and up-regulated the ALP activity as well as the expressions of mineralization-related genes including DSPP, DMP1, ALP, and Nes. Meanwhile, the protein levels of DSP and DMP1 significantly increased in the presence of exogenous Dlx3. Conclusions: Dlx3 is a potent regulator for proliferation and odontoblastic differentiation of hDPCs. (J Endod 2012;38:1504-1510)