1. Absence of K-Ras Reduces Proliferation and Migration But Increases Extracellular Matrix Synthesis in Fibroblasts.
- Author
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Muñoz-Félix JM, Fuentes-Calvo I, Cuesta C, Eleno N, Crespo P, López-Novoa JM, and Martínez-Salgado C
- Subjects
- Animals, Butadienes pharmacology, Cells, Cultured, Extracellular Matrix Proteins metabolism, Fibronectins metabolism, Mice, Mice, Knockout, Nitriles pharmacology, Phosphatidylinositol 3-Kinases metabolism, Phosphorylation, Proto-Oncogene Proteins p21(ras) deficiency, Transforming Growth Factor beta1 metabolism, Cell Movement drug effects, Cell Proliferation drug effects, Extracellular Matrix metabolism, Fibroblasts metabolism, Proto-Oncogene Proteins p21(ras) metabolism, Signal Transduction
- 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., (© 2016 Wiley Periodicals, Inc.)
- Published
- 2016
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