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Inhibition of NADPH oxidase 4 activates apoptosis via the AKT/apoptosis signal-regulating kinase 1 pathway in pancreatic cancer PANC-1 cells

Oncogene volume 25, pages 3699–3707 (2006)Cite this article

Abstract

Pancreatic adenocarcinoma is an aggressive human malignancy and is characterized by resistance to apoptosis. Recently, NADPH oxidase (Nox) 4-mediated generation of intracellular reactive oxygen species (ROS) was proposed to confer antiapoptotic activity and thus a growth advantage to pancreatic cancer cells. The signaling mechanism by which Nox4 transmits cell survival signals remains unclear. Here, we show that both a flavoprotein inhibitor, diphenylene iodonium (DPI), and small interfering RNAs designed to target Nox4 mRNA (siNox4RNAs) inhibited superoxide production in PANC-1 pancreatic cancer cells, and depletion of ROS by DPI or siNox4RNAs induced apoptosis. Parallely, DPI treatment and siNox4RNA transfection blocked activation of the cell survival kinase AKT by attenuating phosphorylation of AKT. Furthermore, AKT phosphorylation of apoptosis signal-regulating kinase 1 (ASK1) on Ser-83 was reduced by DPI and siNox4RNAs. When ASK1Ser83Ala (an AKT phosphorylation-defective ASK1 mutant) was introduced into PANC-1 cells, this mutant alone induced apoptosis. But, addition of DPI or co-transfection of siNox4RNA had no additive effect, indicating that the mutant can substitute for these reagents in apoptosis induction. Taken together, these findings suggest that ROS generated by Nox4, at least in part, transmit cell survival signals through the AKT–ASK1 pathway in pancreatic cancer cells and their depletion leads to apoptosis.

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Acknowledgements

We thank Drs M Chao and Y Gotoh for ASK1 and AKT plasmids, respectively. We are grateful to Dr B Goldstein for providing Nox4 antibodies. We thank M Kubodera for technical assistance and Mrs F Ushiyama for assistance in manuscript preparation. This study was supported by Grant-in-Aid in Cancer Research from the Ministry of Science and Culture of Japan (TK).

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Author notes

  1. T Mochizuki, S Furuta and J Mitsushita: These authors contributed equally to this work.

Authors and Affiliations

  1. The Second Department of Internal Medicine, Shinshu University Graduate School of Medicine, Matsumoto, Nagano, Japan

    T Mochizuki & K Kiyosawa

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

    S Furuta, J Mitsushita, W H Shang, M Ito, Y Yokoo & T Kamata

  3. Department of Dermatology, Shinshu University Graduate School of Medicine, Matsumoto, Nagano, Japan

    M Yamaura

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

    S Ishizone

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

    J Nakayama

  6. Biomedical Research Institute, Kureha Chemical Industry Co. Ltd, Hiyakunin-cho, Tokyo, Japan

    A Konagai & K Hirose

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  1. T Mochizuki
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  2. S Furuta
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Correspondence to T Kamata.

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Mochizuki, T., Furuta, S., Mitsushita, J. et al. Inhibition of NADPH oxidase 4 activates apoptosis via the AKT/apoptosis signal-regulating kinase 1 pathway in pancreatic cancer PANC-1 cells. Oncogene 25, 3699–3707 (2006). https://doi.org/10.1038/sj.onc.1209406

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  • DOI: https://doi.org/10.1038/sj.onc.1209406

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