Abstract
Hepatocyte growth factor (HGF), a fibroblast-derived protein that affects the growth, motility and differentiation of epithelial cells, is a mitogen for human melanocytes and has recently been implicated as an important factor for the development and dissemination of melanomas. To better define the possible role of HGF in the multi-step progression from melanocyte to melanoma cell, we used retrovirus-mediated gene transfer to introduce the gene encoding human HGF into normal human melanocytes, thus causing these cells to produce a growth factor they do not normally express and creating a new autocrine loop as sometimes occurs in melanoma cells. Cells were transduced with an efficiency of 6%, and the modified cells synthesized and secreted significant levels of HGF (6.6 ng/10(7) cells per 24 h) in vitro. Cells expressing HGF had a higher rate of proliferation when compared with unmodified cells and formed large, dense, melanin-positive colonies on a plastic surface. Immunostaining showed HGF-positive melanocytes with varying levels of expression, and HGF protein was detected throughout the whole cell. Although proliferation of HGF-expressing melanocytes was enhanced, they failed to form colonies in a soft agar assay. These resul...Continue Reading
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