口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (2): 125-129.DOI: 10.13701/j.cnki.kqyxyj.2018.02.005

• 牙周病学研究 • 上一篇    下一篇

C4orf7在牙周膜细胞成骨分化中的作用及机制

伍云1, 赵艳1,2, 季耀庭1, 夏海滨1,3, 张壁1,2, 王贻宁1,2*   

  1. 1. 武汉大学口腔医学院口腔基础医学省部共建国家重点实验室培训基地和口腔生物医学教育部重点实验室 湖北 武汉 430079;
    2. 武汉大学口腔医院修复科 湖北 武汉 430079;
    3. 武汉大学口腔医院种植科 湖北 武汉 430079;
  • 收稿日期:2017-11-30 出版日期:2018-02-28 发布日期:2018-02-26
  • 通讯作者: 王贻宁,E-mail: wang.yn@whu.edu.cn
  • 作者简介:伍云(1992~ ),女,广西桂林人,硕士在读,主要从事口腔修复学研究工作。
  • 基金资助:
    国家自然科学基金(编号:81400544)

Role of C4orf7 in Osteogenic Differentiation of Human Periodontal Ligament Cells

WU Yun1, ZHAO Yan1,2, JI Yao-ting1, XIA Hai-bing1,3, ZHANG Bi1,2, WANG Yi-ning1,2*   

  1. 1. The State Key Laboratory Breeding Base of Basic Science of Stomatology and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan 430079, China;
    2. Department of Prosthodontics, StomatologyHospital, WuhanUniversity, Wuhan 430079, China;
    3. Department of Implantology, Stomatology Hospital, Wuhan University, Wuhan 430079, China.
  • Received:2017-11-30 Online:2018-02-28 Published:2018-02-26

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摘要： 目的:探讨C4orf7在牙周膜细胞成骨分化中的作用及机制。方法:首先采用免疫组织化学法(IHC)检测C4orf7在几种牙周组织内的表达情况。人牙周膜细胞(PDLCs)体外培养,利用慢病毒载体技术,构建C4orf7过表达细胞(LV-c4orf7)及空载体对照细胞(LV-con)。对两组细胞矿化诱导4 d、7 d、18 d,检测成骨指标碱性磷酸酶(ALP)、Ⅰ型胶原(COL1)、Runt相关转录因子2(RUNX2)及骨钙素(OCN)的表达变化,ALP活性及矿化结节形成情况;最后,利用免疫共沉淀技术检测C4orf7与骨形成蛋白2(BMP2),Ⅰ型胶原(COLⅠ),Ⅲ型胶原(COLⅢ)蛋白间的互作情况。结果:IHC结果显示,C4orf7仅在牙周韧带部位明显表达。成骨诱导后,与对照组相比,过表达组细胞的ALP活性及矿化结节形成明显减弱,成骨指标ALP,COL1,RUNX2,OCN的表达也均受到抑制(P<0.05)。免疫共沉淀表明,C4orf7可与BMP2分子结合,但与COLⅠ和COLⅢ无作用。结论:C4orf7负向调控PDLCs的成骨分化,并可能通过抑制BMP2功能来实现。

关键词: 牙周膜细胞, C4orf7, 慢病毒载体成骨分化

Abstract: Objective: To investigate the effect and potential mechanism of C4orf7 on the osteogenic differentiation of human periodontal ligamentcells (PDLCs). Methods: HE and immunohistochemical (IHC) staining were used to detect the expression of C4orf7 in periodontal tissue. PDLCs were isolated and cultured by enzyme digestion method. Then, PDLCs were transfected with lentiviral vector containing C4orf7 plasmid (LV-c4orf7) or empty vector plasmid (LV-con) and induced inmineralization induction medium for 4,7, and 18 d. Quantification of osteogenic genes (ALP, COL1, RUNX2, and OCN) expression, ALP activity assay, and Alizarin red staining were used to assess the influence of C4orf7 on the osteogenic differentiation of PDLCs. Immunoprecipitation assay was performed to detect the interaction between C4orf7 and BMP2, COLⅠ, and COLⅢ. Results: IHC assay showed that C4orf7 was specifically expressed in periodontal ligament tissue. ALP activity and the mineralization nodules formation were attenuated by C4orf7 overexpression. The mRNA expression levels of osteogenic genes (ALP, COL1, RUNX2, and OCN) were all downregulated in C4orf7 transfected cells. Immunoprecipitation assay indicated that C4orf7 could bind with BMP2 while had no interaction with COLⅠ and COL Ⅲ. Conclusion: C4orf7 may have negative effect on osteogenic differentiation of human PDLCs by interaction with BMP2.

Key words: Periodontal ligament cell, C4orf7, Lentiviral vector transfection, Osteogenic differentiation

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