中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (15): 2769-2772.doi: 10.3969/j.issn.1673-8225.2010.15.026

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

胶原三维立体培养模型中胚肺成纤维细胞与肺癌GLC-82细胞的相互作用

闫江涛1，2，朱运奎2，李继东2，马国成2，王  舒2，王晓芹2   

  1. 1兰州大学第二临床医学院，甘肃省兰州市 730000；2解放军兰州军区兰州总医院呼吸内科，甘肃省兰州市 730050
  • 出版日期:2010-04-09 发布日期:2010-04-09
  • 通讯作者: 朱运奎，硕士，主任医师，解放军兰州军区兰州总医院呼吸内科，甘肃省兰州市 730050 yunkuizhu@yahoo.com.cn
  • 作者简介:闫江涛★，男，1978年生，河南省郏县人，汉族，兰州大学第二临床医学院在读硕士，医师，主要从事肺癌的基础及临床研究。 yanjt07@lzu.cn
  • 基金资助:

    课题受甘肃省自然科学基金(SZS051-A25-088)资助。

Interaction between lung fibroblast and lung cancer GLC-82 cells in a three-dimensional collagen culture model

Yan Jiang-tao 1,2, Zhu Yun-Kui2, Li Ji-Dong2, Ma Guo-cheng2, Wang Shu2, Wang Xiao-Qin2   

  1. 1 Second Clinical Medical College, Lanzhou University, Lanzhou   730000, Gansu Province, China; 2 Department of Respiratory Medicine, Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • Online:2010-04-09 Published:2010-04-09
  • Contact: Zhu Yun-kui, Master, Chief physician, Department of Respiratory Medicine, Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA, Lanzhou 730050, Gansu Province, China yunkuizhu@yahoo.com.cn
  • About author:Yan Jiang-tao★, Studying for master’s degree, Physician, Second Clinical Medical College, Lanzhou University, Lanzhou 730000, Gansu Province, China; Department of Respiratory Medicine, Lanzhou General Hospital of Lanzhou Military Area Command of Chinese PLA, Lanzhou 730050, Gansu Province, China yanjt07@lzu.cn
  • Supported by:

    the Natural Science Foundation of Gansu Province, No. SZS051-A25-088*

PDF

328

摘要：

背景：组织细胞、炎细胞可能在肿瘤细胞的侵袭、转移中发挥着重要作用，而细胞间的相互作用对基质金属蛋白酶有影响。
目的：观察胚肺成纤维细胞与肺癌GLC-82细胞在胶原三维立体培养条件下，相互作用对基质金属蛋白酶1，2，9表达的影响。
方法：采用三维胶原立体培养模型，按比例混合胶原，4倍DMEM和灭菌水，1倍DMEM的液体胶原，其中胶原终浓度为0.75 g/mL。将GLC-82细胞、胚肺成纤维细胞分别单独培养及GLC-82细胞和胚肺成纤维细胞按5∶1混合培养，待胶原固化后加入1倍DMEM培养基，48 h后收集上清液。Western blot法检测基质金属蛋白酶1表达水平，明胶酶谱法测定基质金属蛋白酶2，9的表达。
结果与结论：胚肺成纤维细胞与肺癌GLC-82细胞混合培养组基质金属蛋白酶1及基质金属蛋白酶2，9分泌量大于单独培养组(P < 0.05)。结果提示，三维立体培养条件下，胚肺成纤维细胞与肺癌GLC-82细胞相互作用能通过上调基质金属蛋白酶1，2，9的表达和活化，促进肺癌侵袭和转移。

关键词: 肺癌GLC-82细胞, 胚肺成纤维细胞, 三维胶原立体培养, 基质金属蛋白酶, 心肺组织工程

Abstract:

BACKGROUND: The histocyte and inflammatory cells played an important role in the invasion and metastasis of lung cancer cells. However, cell interaction influenced matrix metalloproteinase.
OBJECTIVE: To observe the influence of interaction between fetal lung fibroblasts (HFL) and lung cancer cells on matrix metalloproteinase-1, 2, and 9 (MMP-1, 2, 9) expression in three-dimensional co-culture.
METHODS: Human lung cancer cells (GLC-82) and HFL were co-cultivated in three-dimensional collagen gels, which were mixed with collagen, 4 time volume DMEM and sterilizing water, as well as 1 time volume DMEM. The final concentration of collagen was 0.75 g/mL. GLC-82 and HFL were cultured individually, and GLC-82 and HFL were then co-cultured according to the ratio of 5:1. After 48 hours, the supernatant was harvested following adding 1 time volume DMEM. MMP-1 expression was detected by Western blot and MMP-2 and 9 expressions were detected by gelatin zymography.
RESULTS AND CONCLUSION: In three-dimensional co-culture of GLC-82 with HFL, the expressions of MMP-1, MMP-2 and MMP-9 were much higher than that in GLC-82 or in HFL alone (P < 0.05). This suggested that the interaction between HFL and lung cancer cells enhanced the invasion and metastasis of lung cancer through up-regulating the secretion and activation of MMP-1, MMP-2, and MMP-9. 

中图分类号: