title: Gelsolin affects the migratory ability of human colon adenocarcinoma and melanoma cells.
abstract: OBJECTIVE
  Formation of different protrusive structures by migrating cells is driven by actin polymerization at the plasma membrane region. Gelsolin is an actin binding protein controlling the length of actin filaments by its severing and capping activity. The main goal of this study was to determine the effect of gelsolin expression on the migration of human colon adenocarcinoma LS180 and melanoma A375 cells.
  
  
  METHODS
  Colon adenocarcinoma cell line LS180 was stably transfected with plasmid containing human cytoplasmic gelsolin cDNA tagged to enhanced green fluorescence protein (EGFP). Melanoma A375 cells were transfected with siRNAs directed against gelsolin. Real-time PCR and Western blotting were used to determine the level of gelsolin. The ability of actin to inhibit DNase I activity was used to quantify monomeric and total actin level and calculate the state of actin polymerization. Fluorescence confocal microscopy was applied to observe gelsolin and vinculin distribution along with actin cytoskeleton organization.
  
  
  RESULTS
  Increased level of gelsolin expression leads to its accumulation at the submembranous region of the cell accompanied by distinct changes in the state of actin polymerization and an increase in the migration of LS180 cells. In addition, LS180 cells overexpressing gelsolin form podosome-like structures as indicated by vinculin redistribution and its colocalization with gelsolin and actin. Downregulation of gelsolin expression in melanoma A375 cells significantly reduces their migratory potential.
  
  
  CONCLUSIONS
  Our experimental data indicate that alterations in the expression level of gelsolin and its subcellular distribution may be directly responsible for determining migration capacity of human cancer cells.
authors.affiliation: Department of Cell Pathology, Faculty of Biotechnology, University of Wrocław, Poland. mmach@ibmb.uni.wroc.pl
authors.forename: Monika
authors.initials: M
authors.lastname: Litwin
authors.forename: Dorota
authors.initials: D
authors.lastname: Nowak
authors.forename: Antonina Joanna
authors.initials: AJ
authors.lastname: Mazur
authors.forename: Dagmara
authors.initials: D
authors.lastname: Baczyńska
authors.forename: Hans Georg
authors.initials: HG
authors.lastname: Mannherz
authors.forename: Maria
authors.initials: M
authors.lastname: Malicka-Błaszkiewicz
mesh_terms: D000199:Actins / Q000378:metabolism; D000230:Adenocarcinoma / Q000378:metabolism*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement; D003110:Colonic Neoplasms / Q000378:metabolism*; D018260:Gelsolin / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008545:Melanoma / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D058105:Polymerization; D016596:Vinculin / Q000378:metabolism
chemical_list: D000199:Actins; D018260:Gelsolin; D016596:Vinculin