title: Whole Genome Amplification in Genomic Analysis of Single Circulating Tumor Cells.
abstract: Investigation of the genome of organisms is one of the major basics in molecular biology to understand the complex organization of cells. While genomic DNA can easily be isolated from tissues or cell cultures of plant, animal or human origin, DNA extraction from single cells is still challenging. Here, we describe three techniques for the amplification of genomic DNA of fixed single circulating tumor cells (CTC) isolated from blood of cancer patients. This amplification is aimed to increase DNA amounts from those of one cell to yields sufficient for different DNA analyses such as mutational analysis including next-generation sequencing, array-comparative genome hybridization (CGH), and quantitative measurement of gene amplifications. Molecular analysis of CTC as liquid biopsy can be used to identify therapeutic targets in personalized medicine directed, e.g. against human epidermal growth factor receptor 2 (HER2) or epidermal growth factor receptor (EGFR) and to stratify the patients to those therapies.
authors.affiliation: Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, Hamburg, 20246, Germany.
authors.forename: Christin
authors.initials: C
authors.lastname: Gasch
authors.affiliation: Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, Hamburg, 20246, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Pantel
authors.affiliation: Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, Hamburg, 20246, Germany. s.riethdorf@uke.de.
authors.forename: Sabine
authors.initials: S
authors.lastname: Riethdorf
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics; D015894:Genome, Human; D023281:Genomics / Q000379:methods*; D006801:Humans; D009369:Neoplasms / Q000175:diagnosis* / Q000235:genetics*; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D014408:Biomarkers, Tumor
keywords: Circulating tumor cells; Genomic analysis; Linker-adapter PCR; Multiple-strand displacement amplification; Single cell analysis; Whole genome amplification