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Nucleic Acids Research Advance Access published online on March 15, 2008
Nucleic Acids Research, doi:10.1093/nar/gkn101
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

High-resolution array comparative genomic hybridization of single micrometastatic tumor cells

Christine Fuhrmann, Oleg Schmidt-Kittler1, Nikolas H. Stoecklein1, Karina Petat-Dutter1, Christian Vay1, Kerstin Bockler1, Richard Reinhardt2, Thomas Ragg3 and Christoph A. Klein1,*

1Division of Oncogenomics, Department of Pathology, University of Regensburg, Franz-Josef-Strauß-Allee 11, 93053 Regensburg, 2Max-Planck Institute for Molecular Genetics, 14195 Berlin, and 3Quantiom Bioinformatics GmbH & Co. KG, Ringstr. 61, 76356 Weingarten, Germany

*To whom correspondence should be addressed. Tel: +49 941 9446720; Fax: +49 941 9446719; Email: christoph.klein{at}klinik.uni-regensburg.de

Received January 29, 2008. Accepted February 22, 2008.

Only few selected cancer cells drive tumor progression and are responsible for therapy resistance. Their specific genomic characteristics, however, are largely unknown because high-resolution genome analysis is currently limited to DNA pooled from many cells. Here, we describe a protocol for array comparative genomic hybridization (array CGH), which enables the detection of DNA copy number changes in single cells. Combining a PCR-based whole genome amplification method with arrays of highly purified BAC clones we could accurately determine known chromosomal changes such as trisomy 21 in single leukocytes as well as complex genomic imbalances of single cell line cells. In single T47D cells aberrant regions as small as 1–2 Mb were identified in most cases when compared to non-amplified DNA from 106 cells. Most importantly, in single micrometastatic cancer cells isolated from bone marrow of breast cancer patients, we retrieved and confirmed amplifications as small as 4.4 and 5 Mb. Thus, high-resolution genome analysis of single metastatic precursor cells is now possible and may be used for the identification of novel therapy target genes.

Present address: Oleg Schmidt-Kittler, Howard Hughes Medical Institute, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins School of Medicine, Baltimore, Maryland 21231, USA.

Nikolas H. Stoecklein and Christian Vay, Klinik für Allgemein-, Viszeral- und Kinderchirurgie, Universitätsklinikum Düsseldorf, Moorenstr. 5, 40225 Düsseldorf, Germany.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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