Matrilysin mediates extracellular cleavage of E-cadherin from prostate cancer cells: A key mechanism in hepatocyte growth factor/scatter factor-induced cell-cell dissociation and in vitro invasion

Purpose: The current study examined the effects of hepatocyte growth factor /scatter factor (HGF/SF) on cell-cell dissociation, invasion, and its assoc iation with the mediated release of matrix metalloproteinase-7 (Matrilysin) on the extracellular cleavage of E-cadherin in prostate cancer cells. Experimental Design: The effects of HGF/SF on cell-cell dissociation, in vi tro invasion, and on the expression of E-cadherin at both protein and mRNA levels were assessed in cells whose expression of Matrilysin was altered by treatment with antisense oligonucleotide. Results: Incubation with HGF/SF mediated the release of active Matrilysin ( M-r 19,000), resulting in extracellular cleavage of E-cadherin from prostat e cancer cells. This resultant soluble M-r 80,000 fragment of E-cadherin wa s subsequently recognized upon immunoprobing with an anti-E-cadherin antibo dy. Both recombinant human Matrilysin (rh-Matrilysin) and/or HGF/SF increas ed the level of soluble E-cadherin and decreased the level of full-length ( M-r 120,000) E-cadherin as detected by Western blotting. The effects of rh- Matrilysin and HGF/SF were inhibited by an antisense oligonucleotide specif ically directed toward human Matrilysin. In addition, stimulation with eith er rh-Matrilysin or HGF/SF resulted in disruption to the E-cadherin/beta -c atenin complex, as shown by a significant increase (P < 0.05) in both cell scattering and invasion index. Conclusions: Treatment with HGF/SF induced Matrilysin-mediated cleavage to the extracellular domain of E-cadherin, resulting in its dissociation from the cadherin/ catenin complex. This provides a new mechanism in HGF/ SF-ind uced cell scattering, resulting in a switch to a more invasive phenotype in LNCapFGC cells, as demonstrated by in vitro invasion.