口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (2): 134-138.DOI: 10.13701/j.cnki.kqyxyj.2018.02.007

• 牙周病学研究 • 上一篇    下一篇

两种细胞培养方法探究wnt/β-catenin信号通路对EMMPRIN/MMPs的影响

刘学姣, 张珍, 李成章*, 尚姝环   

  1. 武汉大学口腔医学院 湖北 武汉 430079;
  • 收稿日期:2017-10-09 出版日期:2018-02-28 发布日期:2018-02-26
  • 通讯作者: 李成章,E-mail:lcz56@163.com
  • 作者简介:刘学姣(1992~ ),女,山东省淄博市人,硕士在读,主要从事牙周病的宿主因素研究。
  • 基金资助:
    国家自然基金资助(编号:81670995、81371146)

Effect of Wnt/β-catenin Signaling Pathway on Expressions of EMMPRIN/MMPs in Two Cell Culture Models

LIU Xue-jiao, ZHANG Zhen, LI Cheng-zhang*, SHANG Shu-huan   

  1. Hospital of Stomatology Wuhan University, Wuhan 430079, China.
  • Received:2017-10-09 Online:2018-02-28 Published:2018-02-26

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摘要： 目的:用牙龈成纤维细胞(human gingival fibroblast,HGFs)单独培养、HGFs与人永生口腔上皮细胞(human immortalized oral epithelial cell,HIOEC)直接共培养模型探究wnt/β-catenin信号通路对基质金属蛋白酶诱导因子(EMMPRIN)、明胶酶(MMP-2、MMP-9)表达的影响。方法:细胞免疫荧光检测β-catenin的表达。使用wnt/β-catenin信号通路的抑制剂Dickkopf-1(DKK-1)和激活剂wnt3a分别作用于HGFs和共培养模型。蛋白印记法、实时荧光定量PCR、明胶酶谱法检测细胞内和培养基上清中相关指标的表达。结果:β-catenin在HIOEC中表达较HGFs多。当wnt/β-catenin信号通路被抑制时,EMMPRIN、MMP-2、MMP-9的表达增加(P＜0.05),反之则减少(P＜0.05)。共培养模型较HGFs单独培养对DKK-1和wnt3a的反应更敏感。结论:共培养模型是本实验较好的实验方法。激活wnt/β-catenin信号通路可有效抑制EMMPRIN、MMP-2、MMP-9的表达。

关键词: 共培养, wnt/β-catenin信号通路, 基质金属蛋白酶诱导因子, 基质金属蛋白酶

Abstract: Objective: To investigate the effect of wnt/β-catenin signaling pathway on expressions of EMMPRIN, MMP-2, and MMP-9 by individual culture or co-culture human gingival fibroblasts (HGFs)and human immortalized oral epithelial cells (HIOEC). Methods: Immunofluorescence staining was used to determinethe expression of β-catenin. Dickkopf-1(DKK-1) and wnt3a were respectively used as inhibitor and activator of wnt/β-catenin signaling pathway to stimulate HGFs and co-culture model.The expressions of relevant indicators in cells and supernatant were detected by western blot, Real-time PCR, and gelatin zymography. Results: The level ofβ-catenin was much higher in HIOEC than HGFs. When thewnt/β-catenin signaling pathway was suppressed, the levels of EMMPRIN, MMP-2, and MMP-9 increased (P＜0.05). On the contrary, when this signaling pathway was activated, the result was opposite (P＜0.05). Conclusion: Co-culture model is a proper method in this experiment, and the expressions of EMMPRIN, MMP-2, and MMP-9 were effectively decreased by activation of wnt/β-catenin signaling pathway.

Key words: Co-culture, Wnt/β-catenin signaling pathway, EMMPRIN, MMPs

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