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Co-treatment with hepatocyte growth factor and TGF-beta1 enhances migration of HaCaT cells through NADPH oxidase-dependent ROS generation.

Authors
Nam, HJ; Park, YY; Yoon, G; Cho, H; Lee, JH
Citation
Experimental & molecular medicine, 42(4):270-279, 2010
Journal Title
Experimental & molecular medicine
ISSN
1226-36132092-6413
Abstract
Wound healing requires re-epithelialization from the wound margin through keratinocyte proliferation and migration, and some growth factors are known to influence this process. In the present study, we found that the co-treatment with hepatocyte growth factor (HGF) and TGF-beta1 resulted in enhanced migration of HaCaT cells compared with either growth factor alone, and that N-acetylcysteine, an antioxidant agent, was the most effective among several inhibitors tested, suggesting the involvement of reactive oxygen species (ROS). Fluorescence-activated cell sorter analysis using 2,7-dichlorofluorescein diacetate (DCF-DA) dye showed an early (30 min) as well as a late (24 h) increase of ROS after scratch, and the increase was more prominent with the growth factor treatment. Diphenyliodonium (DPI), a potent inhibitor of NADPH oxidase, abolished the increase of ROS at 30 min, followed by the inhibition of migration, but not the late time event. More precisely, gene knockdown by shRNA for either Nox-1 or Nox-4 isozyme of gp91phox subunit of NADPH oxidase abolished both the early time ROS production and migration. However, HaCaT cell migration was not enhanced by treatment with H((2))O((2)). Collectively, co-treatment with HGF and TGF-beta1 enhances keratinocyte migration, accompanied with ROS generation through NADPH oxidase, involving Nox-1 and Nox-4 isozymes.
MeSH terms
Biphenyl Compounds/pharmacologyCell Movement/drug effects*Epithelium/drug effectsEpithelium/enzymologyGene Knockdown TechniquesHepatocyte Growth Factor/pharmacology*HumansHydrogen Peroxide/pharmacologyIsoenzymes/metabolismKeratinocytes/cytology*Keratinocytes/drug effectsKeratinocytes/enzymology*NADPH Oxidase/metabolism*Onium Compounds/pharmacologyPhosphatidylinositol 3-Kinases/metabolismProtein Subunits/metabolismReactive Oxygen Species/metabolism*Signal Transduction/drug effectsTime FactorsTransforming Growth Factor beta1/pharmacology*
DOI
10.3858/emm.2010.42.4.026
PMID
20177149
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Biochemistry & Molecular Biology
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