title: H-ras transformation sensitizes volume-activated anion channels and increases migratory activity of NIH3T3 fibroblasts.
abstract: The expression of the H-ras oncogene increases the migratory activity of many cell types and thereby contributes to the metastatic behavior of tumor cells. Other studies point to an involvement of volume-activated anion channels (VRAC) in (tumor) cell migration. In this paper, we tested whether VRACs are required for the stimulation of cell migration upon expression of the H-ras oncogene. We compared VRAC activation and migration of wild-type and H-ras-transformed NIH3T3 fibroblasts by means of patch-clamp techniques and time-lapse video microscopy. Both cell types achieve the same degree of VRAC activation upon maximal stimulation, induced by reducing extracellular osmolarity from 300 to 190 mOsm/l. However, upon physiologically relevant reductions in extracellular osmolarity (275 mOsm/l), the level of VRAC activation is almost three times higher in H-ras-transformed compared to wild-type fibroblasts. This increase in VRAC sensitivity is accompanied by increased migratory activity of H-ras fibroblasts. Moreover, the high-affinity VRAC blocker NS3728 inhibits migration of H-ras fibroblasts dose-dependently by up to about 60%, whereas migration of wild-type fibroblasts is reduced by only about 35%. Consistent with higher VRAC activity in H-ras than in wild-type fibroblasts, more VRAC blocker is needed to achieve a comparable degree of inhibition of migration. We suggest that H-ras modulates the volume set point of VRAC and thus facilitates transient changes of cell volume required for faster cell migration.
authors.affiliation: Department of Molecular Biology, University of Copenhagen, 13 Universitetsparken, 2100 Copenhagen, OE, Denmark.
authors.forename: Linda
authors.initials: L
authors.lastname: Schneider
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Klausen
authors.forename: Christian
authors.initials: C
authors.lastname: Stock
authors.forename: Sabine
authors.initials: S
authors.lastname: Mally
authors.forename: Søren T
authors.initials: ST
authors.lastname: Christensen
authors.forename: Stine Falsig
authors.initials: SF
authors.lastname: Pedersen
authors.forename: Else K
authors.initials: EK
authors.lastname: Hoffmann
authors.forename: Albrecht
authors.initials: A
authors.lastname: Schwab
mesh_terms: D000818:Animals; D002224:Carbanilides; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D005347:Fibroblasts / Q000187:drug effects; D011905:Genes, ras / Q000235:genetics / Q000502:physiology*; D015640:Ion Channel Gating / Q000235:genetics / Q000502:physiology*; D051379:Mice; D041681:NIH 3T3 Cells; D009994:Osmolar Concentration; D018408:Patch-Clamp Techniques; D014170:Transformation, Genetic / Q000235:genetics / Q000502:physiology; D014508:Urea / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D002224:Carbanilides; D014508:Urea; C517730:NS-3728