title: MicroRNA signature characterizes primary tumors that metastasize in an esophageal adenocarcinoma rat model.
abstract: OBJECTIVE
  To establish a miRNA signature for metastasis in an animal model of esophageal adenocarcinoma (EAC).
  
  
  BACKGROUND
  The incidence of esophageal adenocarcinoma (EAC) has dramatically increased and esophageal cancer is now the sixth leading cause of cancer deaths worldwide. Mortality rates remain high among patients with advanced stage disease and esophagectomy is associated with high complication rates. Hence, early identification of potentially metastatic disease would better guide treatment strategies.
  
  
  METHODS
  The modified Levrat's surgery was performed to induce EAC in Sprague-Dawley rats. Primary EAC and distant metastatic sites were confirmed via histology and immunofluorescence. miRNA profiling was performed on primary tumors with or without metastasis. A unique subset of miRNAs expressed in primary tumors and metastases was identified with Ingenuity Pathway Analysis (IPA) along with upstream and downstream targets. miRNA-linked gene expression analysis was performed on a secondary cohort of metastasis positive (n=5) and metastasis negative (n=28) primary tumors.
  
  
  RESULTS
  The epithelial origin of distant metastasis was established by IF using villin (VIL1) and mucin 5AC (MUC5AC) antibodies. miRNome analysis identified four down-regulated miRNAs in metastasis positive primary tumors compared to metastasis negative tumors: miR-92a-3p (p=0.0001), miR-141-3p (p=0.0022), miR-451-1a (p=0.0181) and miR133a-3p (p=0.0304). Six target genes identified in the top scoring networks by IPA were validated as significantly, differentially expressed in metastasis positive primary tumors: Ago2, Akt1, Kras, Bcl2L11, CDKN1B and Zeb2.
  
  
  CONCLUSIONS
  In vivo metastasis was confirmed in the modified Levrat's model. Analysis of the primary tumor identified a distinctive miRNA signature for primary tumors that metastasized.
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Ali H
authors.initials: AH
authors.lastname: Zaidi
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Lindsey T
authors.initials: LT
authors.lastname: Saldin
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Lori A
authors.initials: LA
authors.lastname: Kelly
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Linda
authors.initials: L
authors.lastname: Bergal
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Londono
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Juliann E
authors.initials: JE
authors.lastname: Kosovec
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Komatsu
authors.affiliation: Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Pashtoon M
authors.initials: PM
authors.lastname: Kasi
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Amit A
authors.initials: AA
authors.lastname: Shetty
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Keane
authors.affiliation: Division of Gastroenterology, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Shyam J
authors.initials: SJ
authors.lastname: Thakkar
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Luai
authors.initials: L
authors.lastname: Huleihel
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Landreneau
authors.affiliation: McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Stephen F
authors.initials: SF
authors.lastname: Badylak
authors.affiliation: Esophageal and Lung Institute, Allegheny Health Network, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Blair A
authors.initials: BA
authors.lastname: Jobe
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics*; D000818:Animals; D004195:Disease Models, Animal; D004938:Esophageal Neoplasms / Q000473:pathology*; D020869:Gene Expression Profiling; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D009362:Neoplasm Metastasis / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D035683:MicroRNAs
references.citation: Cancer Res. 2012 Apr 1;72(7):1865-77
references.pmid: 22350417
references.citation: Oncogene. 2008 Mar 13;27(12):1788-93
references.pmid: 17891175
references.citation: Science. 2002 Aug 30;297(5586):1551-5
references.pmid: 12202830
references.citation: Digestion. 2013;87(1):6-11
references.pmid: 23343962
references.citation: Ann Surg. 2013 Jul;258(1):82-8
references.pmid: 23108119
references.citation: Int J Oncol. 2013 Aug;43(2):548-60
references.pmid: 23708087
references.citation: Science. 1999 Nov 26;286(5445):1735-8
references.pmid: 10576740
references.citation: Cell. 2003 Dec 26;115(7):787-98
references.pmid: 14697198
references.citation: J Gastrointest Surg. 2013 Nov 15;:null
references.pmid: 24234242
references.citation: Cancer. 2001 Aug 1;92(3):549-55
references.pmid: 11505399
references.citation: Genes Dev. 2008 Apr 1;22(7):894-907
references.pmid: 18381893
references.citation: Gastroenterology. 2005 May;128(6):1554-66
references.pmid: 15887151
references.citation: Nat Rev Genet. 2009 Oct;10(10):704-14
references.pmid: 19763153
references.citation: Cancer Res. 2008 Aug 1;68(15):6084-91
references.pmid: 18676830
references.citation: Med Mol Morphol. 2006 Dec;39(4):169-75
references.pmid: 17187177
references.citation: Cancer. 2006 Apr 1;106(7):1624-33
references.pmid: 16518827
references.citation: Gut. 2013 Oct;62(10):1406-14
references.pmid: 22917659
references.citation: Surgery. 1992 May;111(5):503-10
references.pmid: 1598670
references.citation: Ann Thorac Surg. 1999 Oct;68(4):1133-6; discussion 1136-7
references.pmid: 10543468
references.citation: Oncogene. 2005 Mar 3;24(10):1718-26
references.pmid: 15608667
references.citation: APMIS. 2010 Apr;118(4):271-6
references.pmid: 20402672
references.citation: Am J Pathol. 2002 May;160(5):1555-60
references.pmid: 12000706
references.citation: Lancet Oncol. 2011 Mar;12(3):296-305
references.pmid: 21109491
references.citation: Carcinogenesis. 2000 Aug;21(8):1587-91
references.pmid: 10910963
references.citation: Cancer Lett. 2014 Aug 1;350(1-2):34-42
references.pmid: 24769072
references.citation: Biochem Biophys Res Commun. 2014 Apr 18;446(4):1197-203
references.pmid: 24680681
references.citation: Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12123-8
references.pmid: 14517352
references.citation: Cell. 2011 Oct 14;147(2):382-95
references.pmid: 22000016
references.citation: Cancer Res. 2009 Sep 15;69(18):7375-84
references.pmid: 19738059
references.citation: Mol Med Rep. 2014 Aug;10(2):652-62
references.pmid: 24888564
references.citation: J Biol Chem. 2014 Feb 14;289(7):4116-25
references.pmid: 24371144
references.citation: J Thorac Cardiovasc Surg. 2008 Feb;135(2):255-60; discussion 260
references.pmid: 18242245
references.citation: Nature. 2005 Oct 13;437(7061):1032-7
references.pmid: 16136080
references.citation: Bioinformatics. 2014 Feb 15;30(4):523-30
references.pmid: 24336805
references.citation: Nat Rev Cancer. 2005 Aug;5(8):591-602
references.pmid: 16056258
references.citation: Langenbecks Arch Surg. 2010 Apr;395(4):347-50
references.pmid: 20300770
references.citation: Expert Opin Biol Ther. 2014 Apr;14(4):483-9
references.pmid: 24506707
references.citation: Biochem Biophys Res Commun. 2011 Feb 11;405(2):153-6
references.pmid: 21232526
references.citation: J Surg Res. 2006 Jul;134(1):1-9
references.pmid: 16488438
references.citation: Nat Biotechnol. 2008 Apr;26(4):400-1
references.pmid: 18392022
references.citation: Mol Carcinog. 2014 May;53(5):360-79
references.pmid: 23192861
references.citation: Front Oncol. 2014 Apr 14;4:64
references.pmid: 24782981
references.citation: Cancer Res. 2010 Jul 15;70(14):6015-25
references.pmid: 20610624
references.citation: Am J Gastroenterol. 1999 Dec;94(12):3427-33
references.pmid: 10606298
references.citation: Cancer Res. 2014 Feb 1;74(3):751-64
references.pmid: 24335959
references.citation: Gastrointest Endosc. 2007 Feb;65(2):185-95
references.pmid: 17258973
references.citation: Oncogene. 2011 Jun 30;30(26):2954-63
references.pmid: 21339740
references.citation: Gastroenterol Clin North Am. 2014 Mar;43(1):1-14
references.pmid: 24503355
references.citation: FEBS Lett. 2014 Aug 19;588(16):2639-52
references.pmid: 24681102
references.citation: Ann Surg Oncol. 2007 Aug;14(8):2406-10
references.pmid: 17534685
references.citation: Mol Med Rep. 2012 Jun;5(6):1401-8
references.pmid: 22446994
references.citation: Cancer. 2013 Jun 1;119(11):1985-93
references.pmid: 23456798