Therapeutic targeting of tumor growth and angiogenesis with a novel anti-S100A4 monoclonal antibody.

S100A4, a member of the S100 calcium-binding protein family secreted by tumor and stromal cells, supports tumorigenesis by stimulating angiogenesis. We demonstrated that S100A4 synergizes with vascular endothelial growth factor (VEGF), via the RAGE receptor, in promoting endothelial cell migration by increasing KDR expression and MMP-9 activity. In vivo overexpression of S100A4 led to a significant increase in tumor growth and vascularization in a human melanoma xenograft M21 model.

A highly efficient electroporation method for the transfection of endothelial cells.

Several approaches have been described for improving transfection efficiencies of endothelial cells but the general observations have indicated that yields of transfected endothelial cells are low, irrespective of the techniques used. Here we describe a transfection procedure performed by means of electroporation, with efficiencies up to 85%, by optimizing several parameters such as the electroporation buffer, number of cells, voltage, capacitance and pulse length. The protocol was applied to three endothelial cell types (HUVECs, HUAECs and HMEC-1) commonly used in 'in vitro' angiogenic assays.