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Identification of amplified genes from SV40 large T antigen-induced rat PNET cell lines by subtractive cDNA analysis and radiation hybrid mapping

Oncogene volume 20, pages 2023–2031 (2001)Cite this article

Abstract

Primitive neuroectodermal tumors (PNETs) such as human medulloblastomas are genetically heterogeneous and therefore poorly understood. In a rat model the SV40 large T antigen was used to induce neoplasms with characteristic features of PNETs. Tumor development requires a latency period of 8–11 months implicating secondary genetic alterations. To identify such secondary alterations we performed comparative analyses of two phenotypically identical PNET-derived cell lines. Indeed, these cell lines displayed distinct high-level amplification sites. Using a combination of subtractive cDNA analysis and radiation hybrid mapping we have now identified genes in the amplicon regions of the two cell lines. Interestingly, one of these genes encodes the rat homolog of a cytosolic branched chain aminotransferase (BCATC) previously shown to be amplified in a mouse teratocarcinoma cell line. We propose that this simple cloning strategy may serve as a powerful tool for the isolation of genes implicated in known chromosomal aberrations in primary tumors and tumor cell lines.

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Acknowledgements

We thank Gerd Landsberg for his engaged and excellent technical assistance. This work was supported by the BONFOR program of the University of Bonn Medical Center (to TA Bayer) and by the Deutsche Forschungsgemeinschaft (SFB 400; to KH Scheidtmann and OD Wiestler). U Preuss was a recipient of a Lise-Meitner-Fellowship from the Ministerium für Wissenschaft und Forschung Nordrhein-Westfalen.

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Author notes

  1. Sascha Weggen

    Present address: Department of Neurosciences, University of California San Diego, La Jolla, California, 92093-0691, CA, USA

  2. Sascha Weggen and Ute Preuss: S Weggen and U Preuss contributed equally

Authors and Affiliations

  1. Department of Neuropathology, University of Bonn Medical Center, Sigmund-Freud-Strasse 25, D-53105 Bonn, Germany

    Sascha Weggen, Torsten Pietsch & Otmar D Wiestler

  2. Department of Psychiatry, University of Bonn Medical Center, Sigmund-Freud-Strasse 25, D-53105 Bonn, Germany

    Thomas A Bayer

  3. Institute of Genetics, University of Bonn, Roemerstrasse 164, D-53117 Bonn, Germany

    Ute Preuss, Ingrid Klawitz & Karl-Heinz Scheidtmann

  4. Psychological Institute, University of Bonn, Roemerstrasse 164, D-53117 Bonn, Germany

    Norbert Hilger

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Weggen, S., Preuss, U., Pietsch, T. et al. Identification of amplified genes from SV40 large T antigen-induced rat PNET cell lines by subtractive cDNA analysis and radiation hybrid mapping. Oncogene 20, 2023–2031 (2001). https://doi.org/10.1038/sj.onc.1204287

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