Altered Interphase Fluorescence in Situ Hybridization Profiles of Chromosomes 4, 8q24, and 9q34 in Pancreatic Ductal Adenocarcinoma Are Associated with.

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Altered Interphase Fluorescence in Situ Hybridization Profiles of Chromosomes 4, 8q24, and 9q34 in Pancreatic Ductal Adenocarcinoma Are Associated with a Poorer Patient Outcome María L. Gutiérrez, Luis Muñoz-Bellvis, María E. Sarasquete, David G. Hernández-Mejía, María del Mar Abad, Oscar Bengoechea, Luis Corchete, María González-González, Jacinto García-García, Marcos Gonzalez, Ines Mota, Alberto Orfao, José M. Sayagues The Journal of Molecular Diagnostics Volume 16, Issue 6, Pages 648-659 (November 2014) DOI: 10.1016/j.jmoldx.2014.06.007 Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Interphase fluorescence in situ hybridization microscopic images of distinct cytogenetic patterns found in pancreatic ductal adenocarcinoma cases for chromosomal alterations with an adverse effect on overall patient survival. Arrowheads show chromosome 4 alterations characterized by polyploidy in association or not with del(4p) (A), gains of an entire chromosome 8 or the 8q24 chromosome region (B), and gains and deletions of the 9q34 chromosomal region (C). LSI, locus-specific identifier probe; SA, spectrum aqua; SG, spectrum green; SO, spectrum orange. The Journal of Molecular Diagnostics 2014 16, 648-659DOI: (10.1016/j.jmoldx.2014.06.007) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 Overall survival curves of pancreatic ductal adenocarcinoma patients classified according to the presence versus absence of cytogenetic copy number changes (CNCs) of chromosome 4 (A), gains of chromosome 8q24 (B), and CNCs of chromosome 9q34 (C) and the overall cytogenetic score established on the basis of the three most informative independent prognostic factors (gains of chromosome 8q and CNCs of both chromosomes 4 and 9q) (D). The prognostic effect on overall survival for the whole series (n = 51; left column) and for patients who underwent complete tumor resection and had negative retroperitoneal surgical margins (R0) (n = 34; right column). P values for overall survival were determined by both the log-rank test and the Wilcoxon test (asterisk). The Journal of Molecular Diagnostics 2014 16, 648-659DOI: (10.1016/j.jmoldx.2014.06.007) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Overall survival curves from an independent external validation cohort of pancreatic ductal adenocarcinoma patients (n = 43)4 grouped according to the presence of cytogenetic copy number changes of chromosomes 8q24 (gains of 8q24) (A), 9q34 (B), and 4 (C) by comparative genomic hybridization arrays and the overall cytogenetic score obtained per case for these three alterations (D). The Journal of Molecular Diagnostics 2014 16, 648-659DOI: (10.1016/j.jmoldx.2014.06.007) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 4 Detailed characterization of the gained 8q24.21 chromosomal segments evaluated by high-density 500K single-nucleotide polymorphism (SNP) arrays in a subset of pancreatic ductal adenocarcinoma patients (n = 20). Commonly gained protein-coding and RNA-coding genes localized in these regions included the POU5F1P, PVT1, hsa-mir-1204, TMEM75, and hsa-mir-1205 genes, the former two genes being localized in the minimal common regions of gain identified at chromosome 8q24. Copy number (CN) gains ≥2.5; high CN gains ≥4. The Journal of Molecular Diagnostics 2014 16, 648-659DOI: (10.1016/j.jmoldx.2014.06.007) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 5 POU5F1B, MYC, and PVT1 transcript (mRNA) expression levels as assessed by quantitative real-time polymerase chain reaction in pancreatic ductal adenocarcinoma (PDAC) tumors (n = 28) classified according to the presence versus absence of chromosome 8q24 gains compared with nontumoral pancreatic tissue (n = 5). MYC mRNA levels from PDAC tumor samples carrying gains of chromosome 8q24 were significantly lower than those found in other PDAC tumors (P = 0.02) and in nontumoral pancreatic tissue. ∗∗∗P < 0.001. The Journal of Molecular Diagnostics 2014 16, 648-659DOI: (10.1016/j.jmoldx.2014.06.007) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions