Reactive oxygen generated by NADPH oxidase 1 (Nox1) contributes to cell invasion by regulating matrix metalloprotease-9 production and cell migration.
A mediating role of the reactive oxygen species-generating enzyme Nox1 has been suggested for Ras oncogene transformation phenotypes including anchorage-independent cell growth, augmented angiogenesis, and tumorigenesis. However, little is known about whether Nox1 signaling regulates cell invasiveness. Here, we report that the cell invasion activity was augmented in K-Ras-transformed normal rat kidney cells and attenuated by transfection of Nox1 small interference RNAs (siRNAs) into the cells. Diphenyleneiodonium (DPI) or Nox1 siRNAs blocked up-regulation of matrix metalloprotease-9 at both protein and mRNA levels in K-Ras-transformed normal rat kidney cells. Furthermore, DPI and Nox1 siRNAs inhibited the activation of IKKalpha kinase and the degradation of IkappaB alpha, suppressing the NFkappaB-dependent matrix metalloprotease-9 promoter activity. Additionally, epidermal growth factor-stimulated migration of CaCO-2 cells was abolished by DPI and Nox1 siRNAs, indicating the requirement of Nox1 activity for the motogenic effect of epidermal growth factor. This Nox1 action was mediated by down-regulation of the Rho activity through the low molecular weight protein-tyrosine phosphatase-p190RhoGAP-dependent mechanism. Taken together, our findings define a mediating role of Nox1-generated reactive oxygen species in cell invasion processes, most notably metalloprotease production and cell motile activity.
Shinohara
Masahiro
M
Department of Molecular Biology and Biochemistry, Shinshu University Graduate School of Medicine, Matsumoto, Nagano 390-8621, Japan.
Adachi
Yoshifumi
Y
Mitsushita
Junji
J
Kuwabara
Mitsuhiro
M
Nagasawa
Atsushi
A
Harada
Saori
S
Furuta
Shuichi
S
Zhang
Yugen
Y
Seheli
Kajla
K
Miyazaki
Hitoshi
H
Kamata
Tohru
T
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D018938:Caco-2 Cells; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D047468:Immunoprecipitation; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000074624:NADPH Oxidase 1; D009861:Onium Compounds / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000502:physiology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014810:Vitamin E / Q000494:pharmacology; D020741:rho GTP-Binding Proteins / Q000378:metabolism
D000975:Antioxidants; D009861:Onium Compounds; D011518:Proto-Oncogene Proteins; D017382:Reactive Oxygen Species; D014810:Vitamin E; D004815:Epidermal Growth Factor; C007517:diphenyleneiodonium; D009247:NADH, NADPH Oxidoreductases; D000074624:NADPH Oxidase 1; C000614712:NOX1 protein, rat; D051550:I-kappa B Kinase; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D020741:rho GTP-Binding Proteins