华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

口腔癌相关成纤维细胞对人工基底膜的降解作用及其机制

宋惠云1, 何昕1, 周红梅2, 李胜富3, 付春华1, 龙丹3   

  1. 1.口腔生物医学工程教育部重点实验室, 四川大学; 2.四川大学华西口腔医院黏膜科; 3.四川大学华西医院卫生部移植工程和移植免疫重点实验室, 四川成都610041
  • 收稿日期:2007-06-25 修回日期:2007-06-25 出版日期:2007-06-20 发布日期:2007-06-20
  • 通讯作者: 周红梅,Tel: 028- 85503480
  • 作者简介:宋惠云(1981-), 女,河南人,硕士
  • 基金资助:

    国家自然科学基金资助项目(30572035);四川省科技攻关资助项目(05SG022- 018- 1)

Investigation of the Degr adation and the mechanisms of Ar tificial Basement Membr ane Co- cultur ed with Or al Car cinoma- associated Fibroblasts

SONG Hui- yun1, HE Xin1, ZHOU Hong- mei2, LI Sheng- fu3, FU Chunhua1, LONG Dan3   

  1. 1. Key. Laboratory of Oral Biomedical Engineering of Ministry of Education, Sichuan University, Chengdu 610041, China; 2. Dept. of Oral Medicine, West China College of Stomatology, Sichuan University, Chengdu 610041, China; 3. Key. Laboratory of Transplantation Engineering and Transplantation Immunity of Ministry of Health, West China Hospital, Sichuan University, Chengdu 610041, China
  • Received:2007-06-25 Revised:2007-06-25 Online:2007-06-20 Published:2007-06-20
  • Contact: ZHOU Hong- mei,Tel: 028- 85503480

摘要:

目的观察口腔癌相关成纤维细胞(CAFs)降解人工基底膜matrigel的能力,并探讨其可能的作用机制。方法matrigel凝胶与CAFs和NFs共同培养24、48、72 h后,收集培养上清液,按照羟脯氨酸检测试剂盒说明测定羟脯氨酸含量;用考马斯亮兰微孔板法行微量蛋白定量;用明胶酶谱分析对基质金属蛋白酶- 2(MMP- 2)、基质金属蛋白酶- 9(MMP- 9)含量进行测定。结果在各时间段,口腔CAFs上清超滤液中羟脯氨酸含量、上清液总蛋白浓度及MMP- 2酶原含量明显高于NFs,且CAFs组表达大量MMP- 2活性酶。口腔CAFs和NFs皆不表达MMP- 9。结论CAFs具有较强的降解细胞外基质的能力,提示其与肿瘤- 宿主微环境中基质重建及肿瘤侵袭有关。

关键词: 癌相关成纤维细胞, 人工基底膜, 降解

Abstract:

Objective To investigate the degradation of artificial basement membrane(matrigel) co- cultured with oral carcinoma- associated fibroblasts (CAFs) and its possible mechanism. Methods CAFs and normal fibroblasts (NFs)were incubated on matrigel for 24, 48, 72 h. Equivalent amounts of conditioned medium were collected and assayed for total protein, hydroxyproline and matrix metalloproteinase- 2(MMP- 2), matrix metalloproteinase- 9(MMP- 9)activity by gelatin zymography. Results Oral CAFs were superior to oral NFs in total protein and hydroxyproline density, CAFs present more pro -MMP - 2 and activated MMP - 2. Conclusion CAFs were superior to NFs in degradation of matrigel. CAFs might play a key role in the reconstitution of extracellular matrix and the progression of tumor.

Key words: carcinoma- associated fibroblasts, artificial basement membrane, degradation