Influence of immunomagnetic enrichment on gene expression of tumor cells
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BioMed Central
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Influence of immunomagnetic enrichment on gene expression of tumor cells Ute Woelfle†, Elisabeth Breit† and Klaus Pantel* Address: Institute of Tumor Biology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany Email: Ute Woelfle - [email protected]; Elisabeth Breit - [email protected]; Klaus Pantel* - [email protected] * Corresponding author †Equal contributors
Published: 16 March 2005 Journal of Translational Medicine 2005, 3:12
doi:10.1186/1479-5876-3-12
Received: 09 December 2004 Accepted: 16 March 2005
This article is available from: http://www.translational-medicine.com/content/3/1/12 © 2005 Woelfle et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: Metastasis is the leading cause of cancer-related death. Bone marrow (BM) is a frequent site for the settlement of disseminated tumor cells which occurs years before overt metastases signal incurability. Methods: Here we describe a new method to assess the initial stage of metastasis development in cancer patients. By immunomagnetic selection with HER2/neu and EpCAM as catcher antigens single disseminated tumor cells can be enriched from BM samples. To examine whether the immunomagnetic enrichment technique may change gene expression in the selected tumor cells, we performed differential expression profiling with the breast cancer cell lines MCF-7 and BT474 as models. The profiles were performed using 1.2 Cancer Arrays (Clontech) containing 1176 cDNAs that can be grouped into different functional categories, such as signal transduction, cell cycle, adhesion, cytoskeleton plasticity, growth factors and others. Results: The reproducibility of the gene expression profiling between duplicate cDNA-array experiments was assessed by two independent experiments with MCF-7 breast cancer cells. Scatter blot analysis revealed a good reproducibility of the cDNA array analysis (i.e. less than 10% difference in the gene expression between the arrays). Subsequent comparative cDNA-array analyses of immunobead-selected and unselected MCF-7 and BT474 cancer cells indicated that the antibody incubation during the immunomagnetic selection procedure did not considerably alter the gene expression profile. Conclusion: The described method offers an excellent tool for the enrichment of micrometastatic tumor cells in BM without largely changing the gene expression pattern of these cells.
Introduction Solid tumors derived from epithelial organs are the main form of cancer in industrialized countries. The first phase of the metastatic development consists of local tumor cell invasion, followed by tumor cell circulation in the blood and homing to secondary distant organs [1,2]. As indicator organ for early systemic dissemination of epithel
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