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Breakpoint characterization of the der(19)t(11;19)(q13;p13) in the ovarian cancer cell line SKOV‐3
Authors:Wiebke Onkes  Regina Fredrik  Francesca Micci  Benjamin J Schönbeck  Jose I Martin‐Subero  Reinhard Ullmann  Felix Hilpert  Karen Bräutigam  Ottmar Janssen  Nicolai Maass  Reiner Siebert  Sverre Heim  Norbert Arnold  Jörg Weimer
Affiliation:1. Department of Obstetrics and Gynaecology, University Medical Center Schleswig‐Holstein, Christian‐Albrechts University, Kiel, Germany;2. Section for Cancer Cytogenetics, Institute for Medical Informatics, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway;3. Centre for Cancer Biomedicine, Institute for Clinical Medicine, Faculty of Medicine, University of Oslo, Norway;4. Molecular Immunology, Institute of Immunology, University Medical Center Schleswig‐Holstein, Kiel, Germany;5. Department of Anatomic Pathology, Pharmacology and Microbiology, University of Barcelona, Barcelona, Spain;6. Department Human Molecular Genetics, Max Planck Institute for Molecular Genetics, 14195 Berlin, Germany;7. Department of Gynecology and Obstetrics, University Medical Center RWTH, Aachen, Germany;8. Institute of Human Genetics, University Hospital Schleswig‐Holstein, Campus Kiel/ University Kiel, Germany
Abstract:About 20% of ovarian carcinomas show alterations of 19p13 and/or 19q13 in the form of added extra material whose origin often is from chromosome 11. Based on earlier spectral karyotype analysis of the ovarian cancer cell line SKOV‐3, which shows an unbalanced translocation der(19)t(11;19), the aim of this study was to determine the precise breakpoints of that derivative chromosome. After rough delimitation of the breakpoints of microdissected derivative chromosomes by array analysis, we designed a matrix of primers spanning 11q13.2 and 19p13.2 detecting multiple amplicons on genomic and cDNA. Sequencing the amplicons, accurate localization of both breakpoints on both chromosomes was possible and we found that exon 14 of HOOK2 from chromosome 19 and exon 2 of ACTN3 from chromosome 11 were fused in the derivative chromosome. The breakpoint in the HOOK2 gene was in an intrinsic triplet of nucleic acids leading to a shift in the ACTN3 reading frame in the derivative chromosome. This frameshift alteration should give rise to an early stop codon causing a loss of function of ACTN3. Signals in two‐dimensional Western blotting exactly match to calculated molecular mass and the isoelectric point of the fusion protein. © 2013 Wiley Periodicals, Inc.
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