Volume 382 of the series Methods in Molecular Biology pp 223-237

Tumor-Stroma Interactions of Metastatic Prostate Cancer Cell Lines

Analyses Using Microarrays
  • Nicolas WernertAffiliated withInstitute of Pathology, University of Bonn
  • , Annette KaminskiAffiliated withInstitute of Pathology, University of Bonn
  • , El-Mustapha HaddoutiAffiliated withInstitute of Pathology, University of Bonn
  • , Jens Claus HahneAffiliated withInstitute of Pathology, University of Bonn

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Tumor-stroma interactions are of great importance not only for the development and progression of primary prostate carcinoma but probably also for the establishment of metastasis. Fibroblasts are an important stromal cell type encountered by metastatic tumor cells at different sites. In previous investigations, we had found that media conditioned by three metastatic prostate cancer cell lines (LNCaP, PC-3, and DU-145) induced cultured nonprostatic fibroblasts to proliferate or to express matrix-metalloproteinase-1 considered important for tumor invasion. Fibroblast-conditioned media in turn stimulate proliferation of DU-145 cells and migration of PC-3 cells. Both tumor cells and fibroblasts secrete VEGF suggesting that not only metastatic but also stromal cells at metastatic sites contribute to the vascularization of metastasis necessary for continuous growth.

In order to better understand the reciprocal tumor-stroma cross-talk in molecular terms we used the mRNA extracted from stimulated and unstimulated neoplastic and fibroblastic stromal cells for cDNA array hybridization using Affymetrix® chips. The three prostate cell lines influenced the fibroblasts nearly in the same manner. In particular proteins involved in cell adhesion, cell-cell contact, and cell cycle regulation were downregulated in stimulated fibroblasts. In contrast, fibroblasts affected every prostate cancer cell line in different ways, which may be because of the different origin of the metastatic prostate cancer cell lines.

Key Words

Microarray prostate cancer metastasis tumor-stroma interactions fibroblasts prostate cancer cell lines RNA extraction