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Characterization of Activated Stroma Cells and Externalized Proteins in 3D Human Breast Cancer Model

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Tumour cells do not exist in isolation but in close contact with host cells comprising the tumour stroma. It is increasingly clear that the cellular and extracellular components of the tumour stroma are integral parts of the tumour environment and contribute considerably to its biological behavior. Stroma cells, such as fibroblasts, adipocytes and endothelial cells, as well as paracrine and autocrine mediators within the tumour microenvironment play critical roles in the biology of malignant cells. It has been shown that fibroblasts contribute to the development of an environment favouring malignant outcome of the tumour growth.

Often cellular co-cultures are used to analyze the tumour microenvironment, however this situation is very far from the in vivo cancer environment. In order to approach a scenario that mimics the actual situation, we have focused on a 3D co-culture setup between cancer and fibroblast cells in Matrigel matrix and this enabled us to observe the direct influence of the two cell types on each other's invasive patterns.

Different mammary epithelial cancer cell lines (MCF7 and a TNF-α sensitive subline, MCF7-s1) as well as fibroblast cell lines (human mammary fibroblast derived, HMF3s, and human skin derived, BJ fibroblasts) were tested in our system.

We found that, in our system the two cancer cell lines exhibited different invasive properties. Moreover fibroblasts of different origin affected the two cancer cell lines differently.

This allowed us to speculate that we are dealing with grades of breast tumours found in human.

We observed reciprocal invasion between cancer cells and fibroblasts when grown in 3D Matrigel co-culture conditions. Furthermore, HMF3s fibroblasts showed site directed invasion towards the cancer cells.

Conditioned media from the 3D co-culture setups (HMF3s and cancer cells) was analyzed and activation of various proteases (e.g. MMP2, -9 and -13) was detected.

In contrast, BJ cells by themselves produce high amounts of activated proteases and could not be further activated by cancer cells. However, up regulation of IL-6 and IL-8 as well as the growth factor GRO (Growth Regulated Oncogene, CXCL1) was observed in BJ cells when in 3D co-culture with MCF7-s1 cells.
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Document Type: Abstract

Affiliations: Kræftens Bekæmpelse, Inst. for biologisk kræftforskning, Strandboulevarden 49, 2100 Købehavn Ø

Publication date: May 1, 2008

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