Mechanisms of invasion induced by hepatocyte growth factor in gallbladder carcinoma cell lines

点击翻译按钮获取中文摘要

Human gallbladder cancers are highly malignant and its prognosis is poor depending on extend of surround tissue invasion. We examined in vitro the invasive activity of four gallbladder cancer cell lines (GB-d1, GB-h3, GB-d2 and FU-GBC-1) in the absence or presence of hepatocyte growth factor (HGF). In type 1 collagen gel culture, HGF stimulated cell proliferation and induced invasive phenotype of arborizing or individual tubule structures in GB-d1, GB-h3 and GB-d2. In Matrigel invasion assay, invasion was also induced in these three cell lines by HGF but not in FU-GBC-1. Cellular motility was however stimulated by HGF in all of the four cell lines at various extent. HGF was produced from primary cultured normal fibroblasts of human gallbladders but not from the gallgaldder cancer cell lines. Zymography for proteolytic enzymes demonstrated high levels of type IV collagenase and urokinase-type plasminogen activator (u-PA) activity in GB-d1, GB-h3 and GB-d2 even in the absence of HGF.In the presence of HGF,the 72K type IV collagenase (MMP-2) activity of GB-h3 and u-PA activities of GB-d1, GB-h3 and GB-d2 were enhanced. In contrast, MMPs and PAs activities of FU-GBC-1 were faint irrespective of HGF addition. Western blot analysis demonstrated higher levels of 190K c-met product (HGF receptor) in GB-d1, GB-h3 and GB-d2 than FU-GBC-1. These results suggest that in addition to the importance of proteolytic potent, the cellular motility induced via HGF/HGF-receptor system is essential for the invasive progression of the gallbladder carcinoma cells.