Ameloblastin regulates cell attachment and proliferation through RhoA and p27.
Ameloblastin regulates cell attachment and proliferation through RhoA and p27.
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成釉素通过 RhoA 和 p27 调节细胞附着和增殖。
DOI：
10.1111/j.1600-0722.2011.00887.x
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发表时间：
2011
影响因子：
1.9
通讯作者：
Luan,Xianghong
中科院分区：
文献类型：
--
作者：
Zhang,Youbin;Zhang,Xu;Lu,Xuanyu;Atsawasuwan,Phimon;Luan,Xianghong
张勇，张旭，陆旭，阿特萨瓦苏万P，滦X.成釉蛋白通过RhoA和p27调节细胞的附着和增殖.欧洲口腔科学杂志，2011；119(增刊.1)：280-285。2011EUR J口腔科学基质黏附蛋白成釉蛋白(AMBN)是骨骼和牙齿矿化基质的独特成分之一。在这里，我们专注于表达AMBN的两种细胞--小鼠牙囊细胞(MDF)和小鼠牙周韧带细胞(MPDL)--以破译AMBN在牙齿、牙周和骨组织发育中的作用。为了检测AMBN的功能，在MDF和mPDL的细胞培养皿中分别加入全长和C端(氨基酸137-407)的重组AMBN蛋白。或者，使用AMBN-小发夹状(Sh)RNA载体对细胞进行瞬时转基因。我们的细胞培养研究表明，包被全长AMBN的培养皿最早在种植后1 h促进mPDL和MDF细胞的附着。为了确定可能有助于AMBN对粘附力影响的潜在中间体，我们评估了AMBN涂层和未涂层对照培养皿中RhoA的表达水平。这些研究表明，AMBN在接种后4 h诱导RhoA的表达，尤其是在mPDL细胞中。培养4 h后，细胞周期抑制因子p27表达上调。此外，外源性AMBN及其C端片段可抑制MDF和mPDL的增殖。最后，用AmBN-shRNA载体瞬时转染MDF和mPDL细胞后，mPDL细胞中p27的表达下调。综上所述，这些数据表明AMBN通过RhoA影响细胞黏附，通过p27影响细胞周期进程。
Zhang Y, Zhang X, Lu X, Atsawasuwan P, Luan X. Ameloblastin regulates cell attachment and proliferation through RhoA and p27.Eur J Oral Sci 2011; 119 (Suppl. 1): 280–285. © 2011 Eur J Oral SciThe matrix adhesion protein ameloblastin (AMBN) is one of the unique components of the mineralizing matrix of bones and teeth. Here we focused on two types of cells expressing AMBN – mouse dental follicle cells (mDF) and mouse periodontal ligament cells (mPDL) – to decipher AMBN function in developing dental, periodontal, and bone tissues. To test AMBN function, cell culture dishes of mDF and mPDL were exposed to either full‐length or C‐terminal (amino acids 137–407) recombinant Ambn protein. Alternatively, cells were subjected to transient transfection using an Ambn– small hairpin (sh) RNA vector. Our cell culture studies documented that dishes coated with full‐length AMBN promoted the attachment of mPDL and mDF cells as early as 1 h after seeding. In order to identify potential intermediaries that might aid the effect of AMBN on adhesion, RhoA expression levels in AMBN‐coated and uncoated control dishes were assessed. These studies indicated that AMBN induced RhoA expression 4 h after seeding, especially in mPDL cells. After 4 h of culture, the cell cycle inhibitor p27 was also up‐regulated. In addition, exogenous AMBN and its C‐terminal fragment reduced the proliferation of mDF and mPDL. Finally, transient transfection of mDF and mPDL cells with the Ambn–shRNA vector resulted in the down‐regulation of p27 in mPDL cells. Together, these data indicate that AMBN affects cell adhesion via RhoA and cell cycle progression through p27.
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