SUMMARY:The cadherin-mediated cell-cell adhesion system plays a critical role in normal development and morphogenesis.Inactivation of this system is thought to be responsible for cancer invasion and metastasis. A human hepatocellular carcinoma (HCC) cell line, KYN-2, was observed to have great potential for intrahepatic metastasis when orthotopically implanted into the liver of SCID mice. In vitro cultures of KYN-2 cells showed that they formed trabecular structures in suspension but lost tight cell-cell adhesion and became scattered when attached to a substratum such as collagen or fibronectin. In response to adhesion to the substratum, subcellular colocalization of E-cadherin and actin filaments were shown to be reduced, and a significant amount of ␣-catenin was dissociated from the E-cadherin-catenin complex in KYN-2 cells. These changes of cell-cell adhesion were blocked by inhibitory monoclonal antibodies against 1 and 5 integrins. We found that c-Src was coimmunoprecipitated with E-cadherin-catenin complex and was tyrosine-dephosphorylated and activated in the adherent cells. The tyrosine dephosphorylation of c-Src was induced by cell adhesion to the substratum and inhibited by addition of inhibitory monoclonal antibodies against 1 and 5 integrins. These findings indicate that integrin-mediated cell-substratum adhesion inhibits cadherin-mediated cell-cell adhesion, possibly through c-Src activation, and suggest that this cross-talk mediates transient inactivation of the cadherin system and plays an important role in intrahepatic metastasis of human HCC. Modulation of this interaction might provide a new approach to prevent metastasis and recurrence of HCC. (Lab Invest 2000, 80:387-394).M etastasis to other parts of the liver through tumor cell dispersal via the portal vein is a common feature of human hepatocellular carcinoma (HCC) (Nakashima, 1976). This intrahepatic metastasis is observed more frequently than extrahepatic metastasis at a relatively early stage of the disease. It is one of the main causes of recurrence within the liver after initial treatment and for the poor prognosis of HCC patients (Nagao et al, 1990;Yuki et al, 1990).Recently we constructed models of intrahepatic metastasis using orthotopic implantation of human HCC cell lines and identified the critical features of HCC cell lines with high metastatic potential (Genda et al, 1999). The highly-metastatic HCC cell lines showed high motility in vitro, mediated by small guanosine triphosphatase Rho and its downstream effector, Rhoassociated kinase, through actin reorganization. Inhibition of cell motility resulted in prevention of intrahepatic metastasis in vivo, indicating that intrahepatic metastasis of HCC is critically regulated by the motility of cancer cells. Rapid and dynamic alterations of the actin cytoskeleton are observed in motile cells; for example, disruption of cortical actin bundles or stress fibers and formation of filopodia or lamellipodia. These structures of actin filaments are linked via cytoplasmic proteins to tr...