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Three-dimensional coculture of endometrial cancer cells and fibroblasts in human placenta derived collagen sponges and expression matrix metalloproteinases in these cells.

Abstract：

OBJECTIVE:Collagen gel constitutes a valuable tool for the study of cell-matrix interactions; however, it is sometimes difficult to use the gel, in which tumor and stromal cells are cocultured, for immunohistochemistry, because it is easily broken during the process of fixation and embedding in paraffin, especially after long-term culture. METHODS:To examine the interaction between endometrial cancer cells and fibroblasts in tumor invasion, we carried out three-dimensional (3-D) coculture of various endometrial cancer cell lines and fibroblasts in human placenta derived collagen sponges and analyzed the expression and localization of matrix metalloproteinases (MMP) and plasminogen activators (PA) in these cells by immunohistochemistry. RESULTS:After 4 weeks of culture on the collagen sponges, endometrial cancer cells composed stratiform or glandular structures on the layer of extracellular matrix, which was composed from fibloblasts and extracellular matrix. Compared to Ishikawa cells, which were rarely invasive, HEC-1A and HEC-1BE and cocultured fibroblasts showed high invasiveness and strong expression of some proteins. In cell line HEC-1BE, MMP-1, -7, and -9, MT1-MMP, tissue inhibitors of metalloproteinases 2, and uPA showed intensive staining in both cancer cells and fibroblasts by immunohistochemistry. HEC-1A cells and cocultured fibroblasts showed expression patterns similar to those of HEC-1BE. CONCLUSION:These results suggested that expression of MMPs and uPA was accelerated in fibroblasts surrounded by cancer cells. We believe that our 3-D coculture system has merit in that the interaction between cancer cells and stromal cells can be visually analyzed by immunohistochemistry and that experiments for a long period, at least 2 weeks, are possible. Furthermore, it is expected that some animal, e.g., nude mouse, experiments can be replaced by experiments using this culture system.

journal_name

Gynecol Oncol

journal_title

Gynecologic oncology

authors

Tanaka R,Saito T,Ashihara K,Nishimura M,Mizumoto H,Kudo R

doi

10.1016/s0090-8258(03)00335-4

subject

Has Abstract

pub_date

2003-08-01 00:00:00

pages

297-304

issue

eissn

0090-8258

issn

1095-6859

pii

S0090825803003354

journal_volume

pub_type

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Three-dimensional coculture of endometrial cancer cells and fibroblasts in human placenta derived collagen sponges and expression matrix metalloproteinases in these cells.