Abstract
Hepatocyte growth factor (HGF) and E-cadherin are important for epithelial morphogenetic events. We examined the effects of HGF on E-cadherin localization and interaction with beta-catenin in polarized Madin-Darby canine kidney (MDCK) cell monolayers grown on filters. Surface biotinylation experiments showed that HGF increases apically accessible E-cadherin. Confocal immunofluorescence microscopy of HGF-treated cells showed localization of E-cadherin at membrane domains contacting the apical compartment and an increase in accessibility of apically applied antibodies to lateral E-cadherin below the tight junction. Coimmunoprecipitation of beta-catenin/E-cadherin complexes showed that the amount of E-cadherin associated with beta-catenin increased during the first 24 h of HGF treatment with a return to baseline values after 48 and 72 h. Metabolic labeling showed that HGF increased the synthetic rate of beta-catenin and the amount of newly synthesized E-cadherin associated with immunoprecipitated beta-catenin, with the peak effect occurring after 12 h of treatment and returning to baseline after 24 h. HGF treatment inhibited transcytosis of immunoglobulin A by the polymeric immunoglobulin receptor. We conclude that HGF treatment of polarized MDCK cells grown on filters decreases cell polarity and alters E-cadherin/beta-catenin interaction and synthesis.
Highlights
Hepatocyte growth factor (HGF)1 is a polypeptide growth factor with pleiotrophic functions which, depending on target cells and tissues and stage of development, can include mitogenesis, cell motility, and the development and regeneration of organs (1)
We examined the effects of HGF on E-cadherin localization and interaction with -catenin in polarized MadinDarby canine kidney (MDCK) cell monolayers grown on filters
To analyze directly the effects of HGF on polarized epithelial cell functions that may be important for epithelial cell rearrangements during HGF-induced morphogenesis and motogenesis, we tested the effects of HGF on MDCK cells cultured on permeable supports
Summary
Hepatocyte growth factor (HGF)1 is a polypeptide growth factor with pleiotrophic functions which, depending on target cells and tissues and stage of development, can include mitogenesis, cell motility, and the development and regeneration of organs (1). We examined the effects of HGF on E-cadherin localization and interaction with -catenin in polarized MadinDarby canine kidney (MDCK) cell monolayers grown on filters. We conclude that HGF treatment of polarized MDCK cells grown on filters decreases cell polarity and alters E-cadherin/-catenin interaction and synthesis.
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