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Involvement of overexpressed wild-type BRAF in the growth of malignant melanoma cell lines

Oncogene volume 23, pages 8796–8804 (2004)Cite this article

Abstract

Comparative genomic hybridization (CGH) using 40 cell lines derived from malignant melanomas (MMs) revealed frequent amplification at 7q33–q34 containing BRAF gene, which often is mutated in MM. We found this gene to be amplified to a remarkable degree in the MM cell lines that exhibited high-level gains at 7q33–q34 in CGH. Among 40 cell lines, the eight lines that revealed neither BRAF nor NRAS mutations showed even higher levels of BRAF mRNA expression than the 32 mutated lines, although DNA amplification at 7q33–q34 was not detected in every lines overexpressing BRAF. MM cells that carried wild-type BRAF and NRAS showed constitutive overexpression of B-Raf protein and phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), even after serum starvation. Not only downregulation of the endogenously overexpressed wild-type B-Raf by antisense oligonucleotide but also a treatment with an inhibitor of mitogen-activated protein kinase kinase (MAPKK, MEK) reduced phosphorylated ERK1/2 and cell growth, whereas the exogenously expressed wild-type B-Raf promoted cell growth in MM cells. Our results provide the evidence that overexpression of wild-type B-Raf, in part but not always as a result of gene amplification, is one of the mechanisms underlying constitutive activation of the MAPK pathway that stimulates growth of MM cells.

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Acknowledgements

We are grateful to Professor Yusuke Nakamura (Human Genome Center, The Institute of Medical Science, The University of Tokyo), Professor Steven A Rosenberg (Surgery Branch, National Cancer Institute, NIH), Professor Takehisa Iwai (Vascular and Applied Surgery, Tokyo Medical and Dental University), and Professor Kenichi Sugihara (Surgical Oncology, Tokyo Medical and Dental University) for their continuous encouragement throughout this work. We thank Drs Takashi Inozume and Yasufumi Goto for their assistance for maintenance of cell lines, and Ai Watanabe and Yuri Yoshinaga for technical assistance. This work was supported by grants-in-aid for Scientific Research (B) and Scientific Research on Priority Areas (C) (to J Inazawa and I Imoto) and a Center of Excellence Program for Research on Molecular Destruction and Reconstruction of Tooth and Bone (to J Inazawa) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and Core Research for Evolutional Science and Technology of the Japan Science and Technology Corporation (to J Inazawa).

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Authors and Affiliations

  1. Department of Molecular Cytogenetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan

    Hideaki Tanami, Issei Imoto, Akira Hirasawa, Yasuhiro Yuki, Itaru Sonoda, Jun Inoue, Kohichiro Yasui, Akiko Misawa-Furihata & Johji Inazawa

  2. Core Research for Evolutional Science and Technology of Japan Science and Technology Corporation, Saitama, Japan

    Issei Imoto, Jun Inoue, Kohichiro Yasui, Akiko Misawa-Furihata & Johji Inazawa

  3. Division of Cellular Signaling, Institute for Advanced Medical Research, Keio University, School of Medicine, Tokyo, Japan

    Yutaka Kawakami

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Correspondence to Johji Inazawa.

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Tanami, H., Imoto, I., Hirasawa, A. et al. Involvement of overexpressed wild-type BRAF in the growth of malignant melanoma cell lines. Oncogene 23, 8796–8804 (2004). https://doi.org/10.1038/sj.onc.1208152

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