口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (9): 952-955.DOI: 10.13701/j.cnki.kqyxyj.2018.09.008

• 口腔肿瘤学研究 • 上一篇    下一篇

A结构域阳性纤维连接蛋白促进破骨细胞形成及根尖囊肿骨质破坏

陈雅文1,王海丞2*,董伟杰1   

  1. 1. 嘉兴市第一医院口腔科 浙江 嘉兴 314001;
    2. 同济大学,同济大学附属口腔医学院,上海牙组织修复与再生工程技术研究中心 上海 200072
  • 收稿日期:2018-03-17 出版日期:2018-09-28 发布日期:2018-09-25
  • 通讯作者: 王海丞,E-mail:haichengwang@sina.com
  • 作者简介:陈雅文(1987~ ),女,浙江人,硕士,主治医师,主要从事口腔正畸学临床及基础研究。
  • 基金资助:
    国家自然科学基金青年基金项目(编号: 81600836); 嘉兴市科技计划项目(编号:2016BY28004)

Extra Domain A Positive Fibronectin Promotes Osteoclast Formation and Bone Destruction of Radicular Cysts.

CHEN Ya-wen1,WANG Hai-Cheng2*, DONG Wei-jie1.   

  1. 1.Department of Stomtology, the First Hospital of Jiaxing, Jiaxing 314001, China;
    2. Department of Pathology, School & Hospital of Stomatology, Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, Shanghai 200072, China.
  • Received:2018-03-17 Online:2018-09-28 Published:2018-09-25

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摘要： 目的: 检测纤维连接蛋白(fibronectin, FN)可变剪接亚型蛋白在根尖囊肿纤维囊壁中的表达,研究每种亚型在体外诱导形成破骨细胞的能力,探讨FN的可变剪接亚型与根尖囊肿病变的进展和骨破坏的关系。方法: 选择8例经病理诊断确诊为根尖囊肿患者手术标本,采用免疫组织化学方法,研究每种FN亚型在病变组织中表达强度,分离成纤维细胞,采用反转录-聚合酶链反应(reverse transcription-poymerase chain reaction, RT-PCR)方法检测总FN及其3种亚型(EDA+FN、EDB+FN、CS1-FN)mRNA的表达水平,并采用条件培养基体外诱导破骨细胞分化。根据每种FN亚型的相对表达情况,研究FN亚型与破骨细胞分化之间的关系。结果: 在FN产生的亚型中,含蛋白结构域A的纤维连接蛋白(EDA+FN)在纤维囊壁中染色强度高于EDB+FN和CS1-FN。囊壁成纤维细胞中,EDA + FN的mRNA水平显著高于EDB+FN (P=0.007) 和 CS1-FN (P=0.003),且EDA+FN/总FN值在成纤维细胞的所有亚型中都是最高的(EDB+FN/总FN,P<0.001 ;CS1-FN/总FN,P<0.001)。体外诱导破骨细胞形成结果显示,EDA+FN/总FN与破骨细胞生成数量呈正相关(R=0.776, P=0.024)。结论: 根尖囊肿囊壁成纤维细胞产生各种FN亚型, EDA+FN为主要类型,且EDA+FN/总FN与成纤维细胞诱导破骨细胞效率呈正相关。提示成纤维细胞产生的EDA+FN有利于导致微环境中破骨细胞形成,促进根尖囊肿的骨破坏。

关键词: 纤维连接蛋白, 蛋白结构域A, 破骨细胞, 根尖囊肿

Abstract: Objective: To detect the expression of the isoforms of fibronectin (FN) variable splicing subtype protein in radicular cysts, and to study the ability of each subtype in inducing osteoclasts in vitro, and to analyze the association of these isoforms with osteoclastogenesis. Methods: Specimens from 8 patients with radicular cysts were selected. Immunohistochemistry was used to study the expression of each isoforms in the fibrous capsule. Fibroblasts were isolated from surgical samples, the mRNA levels of each isoform and total FN were detected by RT-PCR, and the conditioned medium was collected to induce the osteoclasts in vitro. The association between FN isoforms and osteoclastogenesis was analyzed. Results: In the fibrous capsule of radicular cysts, FN containing the extra domain A (EDA+FN) was stained more intense than EDB+FN and CS1-FN. Consistently, the mRNA level of EDA+FN was also significantly higher than that of EDB+FN (P=0.007) and CS1-FN (P=0.003) in the fibroblasts. The ratio of EDA+FN/total FN was also the highest among three isoforms (EDB+FN/total FN,P<0.001 and CS1-FN/total FN, P<0.001). Only EDA+FN/total FN was positively associated with the Trap+MNC numbers induced by the conditioned medium (n=8, R=0.776, P=0.024). Conclusion: The fibroblasts in the fibrous capsule of radicular cysts generate various FN isoforms, in which EDA+FN consisted of the majority and positively associated the oste-
oclastogenesis induced by fibroblasts. It is suggested that the fibroblasts within the stroma made the microenvironment favourable to the bone destruction of radicular cysts.

Key words: Fibronectin Extra domain, A Alternative splicing , Osteoclasts , Radicular cyst