caa a22 4500 606190651 CHVBK 20210128100856.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s10577-015-9474-8 doi (NATIONALLICENCE)springer-10.1007/s10577-015-9474-8 A novel canine kidney cell line model for the evaluation of neoplastic development: karyotype evolution associated with spontaneous immortalization and tumorigenicity [Elektronische Daten] [R. Omeir, R. Thomas, B. Teferedegne, C. Williams, G. Foseh, J. Macauley, L. Brinster, J. Beren, K. Peden, M. Breen, A. Lewis Jr] The molecular mechanisms underlying spontaneous neoplastic transformation in cultured mammalian cells remain poorly understood, confounding recognition of parallels with the biology of naturally occurring cancer. The broad use of tumorigenic canine cell lines as research tools, coupled with the accumulation of cytogenomic data from naturally occurring canine cancers, makes the domestic dog an ideal system in which to investigate these relationships. We developed a canine kidney cell line, CKB1-3T7, which allows prospective examination of the onset of spontaneous immortalization and tumorigenicity. We documented the accumulation of cytogenomic aberrations in CKB1-3T7 over 24months in continuous culture. The majority of aberrations emerged in parallel with key phenotypic changes in cell morphology, growth kinetics, and tumor incidence and latency. Focal deletion of CDKN2A/B emerged first, preceding the onset and progression of tumorigenic potential, and progressed to a homozygous deletion across the cell population during extended culture. Interestingly, CKB1-3T7 demonstrated a tumorigenic phenotype in vivo prior to exhibiting loss of contact inhibition in vitro. We also performed the first genome-wide characterization of the canine tumorigenic cell line MDCK, which also exhibited CDKN2A/B deletion. MDCK and CKB1-3T7 cells shared several additional aberrations that we have reported previously as being highly recurrent in spontaneous canine cancers, many of which, as with CDKN2A/B deletion, are evolutionarily conserved in their human counterparts. The conservation of these molecular events across multiple species, in vitro and in vivo, despite their contrasting karyotypic architecture, is a powerful indicator of a common mechanism underlying emerging neoplastic activity. Through integrated cytogenomic and phenotypic characterization of serial passages of CKB1-3T7 from initiation to development of a tumorigenic phenotype, we present a robust and readily accessible model (to be made available through the American Type Culture Collection) of spontaneous neoplastic transformation that overcomes many of the limitations of earlier studies. The Author(s), 2015 Neoplastic transformation nationallicence Tumorigenicity nationallicence Canine chromosomes nationallicence Comparative genomic hybridization (CGH) nationallicence Fluorescence in situ hybridization (FISH) nationallicence Madin-Darby canine kidney (MDCK) cell line nationallicence CKB1-3T7 cell line nationallicence AGMK : African green monkey kidney nationallicence ATCC : American Type Culture Collection nationallicence BAC : Bacterial artificial chromosome nationallicence CFA : Canis familiaris nationallicence CNA : Copy number aberration(s) nationallicence CDKN : Cyclin-dependent kinase inhibitor nationallicence CHORI : Children's Hospital Oakland Research Institute nationallicence DMEM : Dulbecco's modified Eagle's medium nationallicence DT : Doubling time nationallicence FBS : Fetal bovine serum nationallicence FISH : Fluorescence in situ hybridization nationallicence IACUC : Institutional Animal Care and Use Committee nationallicence iPS cells : Induced pluripotent stem cells nationallicence MDCK : Madin-Darby canine kidney nationallicence NIH : National Institutes of Health nationallicence oaCGH : Oligonucleotide-array comparative genomic hybridization nationallicence p : Passage nationallicence PAR : Pseudoautosomal region nationallicence PBS : Phosphate-buffered saline nationallicence PDL : Population doubling level nationallicence RPCI : Roswell Park Cancer Institute nationallicence SLP : Single-locus probe nationallicence UCSC : University of California Santa Cruz nationallicence UPD : Uniparental disomy nationallicence Omeir R. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut Thomas R. Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, 27607, Raleigh, NC, USA aut Teferedegne B. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut Williams C. Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, 27607, Raleigh, NC, USA aut Foseh G. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut Macauley J. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut Brinster L. Division of Veterinary Resources, National Institutes of Health, 20892, Bethesda, MD, USA aut Beren J. Office of Counter-Terrorism and Emergency Coordination, Center for Drug Evaluation and Research, Food and Drug Administration, 20993, Silver Spring, MD, USA aut Peden K. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut Breen M. Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, 27607, Raleigh, NC, USA aut Lewis Jr A. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut Chromosome Research Springer Netherlands 23/4(2015-12-01), 663-680 0967-3849 23:4<663 2015 23 10577 https://doi.org/10.1007/s10577-015-9474-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10577-015-9474-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Omeir R. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Thomas R. Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, 27607, Raleigh, NC, USA aut NATIONALLICENCE 700 1- Teferedegne B. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Williams C. Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, 27607, Raleigh, NC, USA aut NATIONALLICENCE 700 1- Foseh G. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Macauley J. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Brinster L. Division of Veterinary Resources, National Institutes of Health, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Beren J. Office of Counter-Terrorism and Emergency Coordination, Center for Drug Evaluation and Research, Food and Drug Administration, 20993, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Peden K. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut NATIONALLICENCE 700 1- Breen M. Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, 27607, Raleigh, NC, USA aut NATIONALLICENCE 700 1- Lewis Jr A. Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Ave, 20993, Silver Spring, MD, USA aut NATIONALLICENCE 773 0- Chromosome Research Springer Netherlands 23/4(2015-12-01), 663-680 0967-3849 23:4<663 2015 23 10577