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NADPH oxidase 1 plays a critical mediating role in oncogenic Ras-induced vascular endothelial growth factor expression

Oncogene volume 27, pages 4724–4732 (2008)Cite this article

Abstract

Reactive oxygen species (ROS)-generating enzyme Nox1 is important in the induction of oncogenic Ras transformation phenotypes, but it is not defined whether Nox1 is involved in Ras-induced upregulation of vascular endothelial growth factor (VEGF), a potent stimulator of tumor angiogenesis. Here we describe that ablation of the Nox1 activity by Nox1 small-interference RNAs (siRNAs) or diphenylene iodonium (DPI) inhibited synthesis of both VEGF proteins and VEGF mRNAs in K-Ras transformed normal rat kidney (KNRK) cells. Nox1siRNAs and DPI suppressed extracellular signal-regulated kinase (ERK)-dependent phosphorylation of a transcription factor Sp1 and Sp1 binding to a VEGF promoter. Furthermore, tumors derived from Nox1siRNA-transfected KNRK cells markedly decreased neovascularization. The Nox1 activity was required for VEGF production in human colon cancer CaCO-2 cells, as in the case of KNRK cells. However, since overexpression of Nox1 in normal rat kidney cells failed to induce VEGF, the Nox1 activity alone was not sufficient to upregulate VEGF expression, which suggests that unlike the previously proposed model, Nox1 may act in concert with other effectors integrated into the Ras network. We propose that Nox1 mediates oncogenic Ras-induced upregulation of VEGF and angiogenesis by activating Sp1 through Ras-ERK-dependent phosphorylation of Sp1.

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Acknowledgements

This work was supported by a grant on Cancer Research in Applied Areas from the Ministry of Science and Culture of Japan (TK). We thank Dr M Shibuya, Dr T Finkel, Dr J Downward and Dr H Sobue for VEGF, catalase, RasVal12 and Cu/ZnSOD plasmids, respectively. We also thank Mrs F Ushiyama for assistance in manuscript preparation.

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Author notes

  1. D Komatsu and M Kato: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, Shinshu University Graduate School of Medicine, Matsumoto, Nagano, Japan

    D Komatsu, M Kato & T Kamata

  2. Department of Surgery, Shinshu University Graduate School of Medicine, Nagano, Japan

    D Komatsu & S Miyagawa

  3. Institute of Organ Transplantation, Reconstructive Medicine and Tissue Engineering, Shinshu University Graduate School of Medicine, Matsumoto, Nagano, Japan

    J Nakayama

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  1. D Komatsu
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  4. S Miyagawa
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  5. T Kamata
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Correspondence to T Kamata.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Komatsu, D., Kato, M., Nakayama, J. et al. NADPH oxidase 1 plays a critical mediating role in oncogenic Ras-induced vascular endothelial growth factor expression. Oncogene 27, 4724–4732 (2008). https://doi.org/10.1038/onc.2008.102

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