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KMID : 0620920100420040270
Experimental & Molecular Medicine
2010 Volume.42 No. 4 p.270 ~ p.279
Co-treatment with hepatocyte growth factor and TGF-¥â1 enhances migration of HaCaT cells through NADPH oxidase-dependent ROS generation
Nam Hyun-Ja

Park Yun-Yeon
Yoon Gye-Soon
Cho Hye-Seong
Lee Jae-Ho
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 hapatocyte growth factor (HGF) and TGF-¥â1 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 H2O2. Collectively, co-treatment with HGF and TGF-¥â1 enhances keratinocyte migration, accompanied with ROS generation through NADPH oxidase, involving Nox-1 and Nox-4 isozymes.
KEYWORD
cell movement, hepatocyte growth factor, keratinocytes, NADPH oxidase, reactive oxygen species, transforming growth factor ¥â1
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