Versican gene: Regulation by the β-catenin signaling pathway plays a significant role in dermal papilla cell aggregative growth
Versican gene: Regulation by the β-catenin signaling pathway plays a significant role in dermal papilla cell aggregative growth
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DOI：
10.1016/j.jdermsci.2012.09.011
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发表时间：
2012-12-01
影响因子：
4.6
通讯作者：
Lu, Yuangang
中科院分区：
文献类型：
--
作者：
Yang, Yadong;Li, Yuanchao;Lu, Yuangang
背景资料：毛乳头细胞（Dermal papilla cells，DPC）在体外培养中表现出多层聚集生长的特性，与毛囊的诱导形成密切相关，并与毛囊的发育和周期调控有关。Versican是一种大的硫酸软骨素蛋白聚糖，是细胞外基质的主要成分之一，在毛囊形成中起着重要作用。目的：研究多功能蛋白聚糖（versican）和β-catenin在毛原细胞（DPC）聚集生长中的作用，探讨多功能蛋白聚糖（versican）基因通过TCF-4/β-catenin信号通路的表达调控机制。本实验首先培养不同代次的DPC，通过RT-PCR和Western blot检测不同代次DPC中β-catenin和versican表达的变化。然后敲除多功能蛋白聚糖和β-catenin基因，通过EMSA和ChIP检测，评价并验证TCF-4/β-catenin与多功能蛋白聚糖基因启动子区TOP元件的结合能力，最后观察Wnt/β-catenin通路抑制对DPC聚集生长的影响。随着传代次数的增加，DPC的聚集性逐渐丧失，多功能蛋白聚糖mRNA和蛋白水平逐渐下降，而β-连环蛋白下降趋势不明显。β-catenin siRNA转染后，β-catenin和versican的mRNA水平同时下降。随着DPC传代次数的增加，不同代次的DPC与多功能蛋白聚糖启动子的结合能力逐渐减弱。结论：Wnt/beta-catenin信号转导系统通过作用于versican基因上游启动子，改变versican的表达，从而调控DPC的聚集生长。(C)2012年日本皮肤病研究学会。由Elsevier爱尔兰有限公司出版。保留所有权利。
Background: Dermal papilla cells (DPCs), which exhibit a multilayer aggregative growth character in in vitro culture, are closely related to induction of hair follicles (HFs) formation, and are associated with the development and cycle regulation of HFs. Versican, a large chondroitin sulfate proteoglycan and one of the major components of the extracellular matrix, plays an essential role in hair follicle formation. And also the Wnt/beta-catenin signaling pathway performs a crutial function in induction during hair follicle growth and embryogenesis.Objective: To characterize the role of versican and beta-catenin in regulating DPCs aggregative growth, and to explore the versican gene expression regulation mechanism by TCF-4/beta-catenin signaling pathway.Methods: We first cultured DPCs at different passages, and detected the change in beta-catenin and versican expression in DPCs of various passages by RT-PCR and Western blot. Then we knockdowned the versican and beta-catenin gene, evaluated and verificated the binding capability of TCF-4/beta-catenin to TOP elements in versican gene promoter region at varied passage DPCs by EMSA and ChIP Assay, finally observed the effect of Wnt/beta-catenin pathway inhibition on DPC aggregative growth.Results: With the increase of passage, DPCs lost the aggregative property, the versican mRNA and protein level in DPCs was on a gradual decline, while not significant declining tendency of beta-catenin. The mRNA of both beta-catenin and versican reduced simultaneously after beta-catenin siRNA transfection. The binding ability of TCF-4/beta-catenin of varied-passage DPCs to cultured versican promoters diminished with the increase of DPC passages. And versican inhibition or Wnt/beta-catenin pathway blocking could both produce considerable effect on the aggregative growth of low-passage DPCs.Conclusion: Wnt/beta-catenin signal transducting system regulates DPC aggregative growth through modifying versican expression by means of acting on the versican gene upstream promoter. (C) 2012 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.