Absence of K-Ras Reduces Proliferation and Migration But Increases Extracellular Matrix Synthesis in Fibroblasts

J Cell Physiol. 2016 Oct;231(10):2224-35. doi: 10.1002/jcp.25340. Epub 2016 Mar 6.

Abstract

The involvement of Ras-GTPases in the development of renal fibrosis has been addressed in the last decade. We have previously shown that H- and N-Ras isoforms participate in the regulation of fibrosis. Herein, we assessed the role of K-Ras in cellular processes involved in the development of fibrosis: proliferation, migration, and extracellular matrix (ECM) proteins synthesis. K-Ras knockout (KO) mouse embryonic fibroblasts (K-ras(-/-) ) stimulated with transforming growth factor-β1 (TGF-β1) exhibited reduced proliferation and impaired mobility than wild-type fibroblasts. Moreover, an increase on ECM production was observed in K-Ras KO fibroblasts in basal conditions. The absence of K-Ras was accompanied by reduced Ras activation and ERK phosphorylation, and increased AKT phosphorylation, but no differences were observed in TGF-β1-induced Smad signaling. The MEK inhibitor U0126 decreased cell proliferation independently of the presence of K-ras but reduced migration and ECM proteins expression only in wild-type fibroblasts, while the PI3K-AKT inhibitor LY294002 decreased cell proliferation, migration, and ECM synthesis in both types of fibroblasts. Thus, our data unveil that K-Ras and its downstream effector pathways distinctively regulate key biological processes in the development of fibrosis. Moreover, we show that K-Ras may be a crucial mediator in TGF-β1-mediated effects in this cell type. J. Cell. Physiol. 231: 2224-2235, 2016. © 2016 Wiley Periodicals, Inc.

MeSH terms

  • Animals
  • Butadienes / pharmacology
  • Cell Movement* / drug effects
  • Cell Proliferation* / drug effects
  • Cells, Cultured
  • Extracellular Matrix / metabolism*
  • Extracellular Matrix Proteins / metabolism
  • Fibroblasts / metabolism*
  • Fibronectins / metabolism
  • Mice
  • Mice, Knockout
  • Nitriles / pharmacology
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphorylation
  • Proto-Oncogene Proteins p21(ras) / deficiency
  • Proto-Oncogene Proteins p21(ras) / metabolism*
  • Signal Transduction*
  • Transforming Growth Factor beta1 / metabolism

Substances

  • Butadienes
  • Extracellular Matrix Proteins
  • Fibronectins
  • Nitriles
  • Transforming Growth Factor beta1
  • U 0126
  • Hras protein, mouse
  • Proto-Oncogene Proteins p21(ras)