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Stress via p53 pathway causes apoptosis by mitochondrial Noxa upregulation in doxorubicin-treated neuroblastoma cells

Oncogene volume 27, pages 741–754 (2008)Cite this article

Abstract

In this study, we employed a panel of cell lines to determine whether p53-dependent cell death in neuroblastoma (NB) cells is caused by apoptotic cellular function, and we further studied the molecular mechanism of apoptosis induced via the p53-dependent pathway. We obtained evidence that a type of p53-dependent stress, doxorubicin (Doxo) administration, causes accumulation of p53 in the nucleus of NB cells and phosphorylation of several serine residues in both Doxo-sensitive and -resistant cell lines. Upregulation of p53-downstream molecules in cells and upregulation of Noxa in the mitochondrial fraction were observed only in Doxo-sensitive NB cells. Significance of Noxa in the Doxo-induced NB cell death was confirmed by Noxa-knockdown experiments. Mitochondrial dysfunction, including cytochrome-c release and membrane potential disregulation, occurred and resulted in the activation of the intrinsic caspase pathway. However, in the Doxo-resistant cells, the accumulation in the nucleus and phosphorylation of p53 did not induce p53-downstream p21Cip1/Waf1 expression and the Noxa upregulation, resulting in the retention of the mitochondrial homeostasis. Taken together, these findings indicate that the p53 pathway seems to play a crucial role in NB cell death by Noxa regulation in mitochondria, and inhibition of the induction of p53-downstream effectors may regulate drug resistance of NB cells.

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Acknowledgements

We are deeply indebted to Professor Kenichi Koike (Department of Pediatrics, Shinshu University School of Medicine) for his excellent advice. We thank Kumiko Sakurai, Yozo Nakazawa, and Jun Miki for their technical assistance, and Daniel Mrozek, Medical English Service Inc, for editorial assistance. This work was supported by grants from the Japanese Ministry of Education, Science, Sports and Culture, Grant-in-Aid for Scientific Research (C) (contract nos: 15591098 and 17591077).

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Authors and Affiliations

  1. Department of Pediatrics, Shinshu University School of Medicine, Matsumoto, Nagano, Japan

    K Kurata, R Yanagisawa & T Kamijo

  2. Division of Biochemistry, Chiba Cancer Center Research Institute, Chuoh-ku, Chiba, Japan

    M Ohira, A Nakagawara & T Kamijo

  3. Department of Biochemistry, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan

    M Kitagawa

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  1. K Kurata
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Correspondence to T Kamijo.

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Kurata, K., Yanagisawa, R., Ohira, M. et al. Stress via p53 pathway causes apoptosis by mitochondrial Noxa upregulation in doxorubicin-treated neuroblastoma cells. Oncogene 27, 741–754 (2008). https://doi.org/10.1038/sj.onc.1210672

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