Induction of invasive growth in a gallbladder cancer cell line by hepatocyte growth factor in vitro.

To study the mechanism of invasion and metastasis of gallbladder cancer cells, we established a cancer cell line, GB-d1, from a metastatic lymphnode of poorly differentiated adenocarcinoma of the gallbladder. GB-d1 cells proliferate well in a dish culture and form small cystic cell clusters in a collagen gel containing 10% fetal bovine serum. A conditioned medium of human embryonic lung fibroblasts (HEL) stimulated the proliferation of GB-d1 cells and induced cell scattering in the dish culture. In the gel culture, the conditioned medium induced a transformation of the spherical clusters to arborizating colonies with tubular projections that mimicked an invasion of cancer cells into the surrounding tissue. Similar results were obtained when 10 ng/ml of human recombinant hepatocyte growth factor (h-rHGF) was added to the culture medium. The proliferative and morphological changes induced by the conditioned medium were inhibited by antiserum against h-HGF. HEL and human gallbladder stromal fibroblast-like cells produced substantial levels of HGF in the culture media, while GB-d1 did not produce any detectable level of HGF. These results suggest that HGF promotes the invasive growth of gallbladder cancer cells in vitro, and it was also suggested that stromal fibroblasts may play an important role in the invasive progression of gallbladder cancer in a paracrine fashion.