Analysis of Primary Breast Tumour Stromal Cells and Their Potential Role in Disease Progression.

It has emerged in recent years that interaction between breast cancer stromal and epithelial cells supports tumour progression through promotion of epithelial cell growth, migration and invasion. Although this is thought to be mediated through both secretion of paracrine factors and cell-cell contact, the precise mechanisms are poorly understood. The aim of this study was to further characterise stromal cells derived from primary breast tumours and potentially elucidate mechanisms through which they exert their action.Following written informed consent, specimens of human breast cancer were harvested from patients undergoing surgery. Cells were isolated from tumour (n=19) and tumour-associated normal (TAN, n=9) regions of breast tissue. Breast tissue obtained from reduction mammoplasty served as normal controls. Following tissue dissociation and digestion, stromal cells were isolated by differential centrifugation. Following culture of stromal cells, RNA was extracted, reverse transcribed and relative quantitative PCR performed using primers targeting Fibroblast Activation Protein (FAP), Transforming Growth Factor β (TGFβ), Transforming Growth Factor β Receptor II (TGFβRII), Matrix Metalloproteinase 3 (MMP3), and Vascular Endothelial Growth Factor A (VEGF A).There was a trend towards increased expression of VEGF A, TGFβRII and MMP3 in tumour compared to normal stromal cells, while the level of TGFβ expression was equivalent in all samples examined. There was a significant positive correlation between expression of the proangiogenic factor VEGF A and TGFβRII (Pearson correlation coefficient r=0.317, p Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4163.