title: MicroRNA-224 negatively regulates p21 expression during late neoplastic progression in inflammatory bowel disease.
abstract: BACKGROUND
  The development of colon cancer represents a major complication in patients with inflammatory bowel disease (IBD). The importance of microRNAs (miRs) in carcinogenesis is becoming clearer because miRs have been implicated in the regulation of cancer-related cellular processes to include apoptosis, differentiation, cell cycle progression, and immune function. In the current study, we sought to identify miR dysregulation specific to progression along the normal-inflammation-cancer axis in colonic specimens from patients with IBD.
  
  
  METHODS
  MiR microarrays and quantitative reverse transcription PCR were used to detect and confirm dysregulated miRs. Receiver operating characteristic curve analysis was applied to evaluate the potential use of miR-224 as a neoplastic disease marker in IBD. For miR-224 target messenger RNA (mRNA) identification, mRNA microarrays were employed in combination with bioinformatic analyses, Western blotting, and luciferase activity measurements.
  
  
  RESULTS
  We identified 30 miRs that were differentially expressed between chronically inflamed mucosae and cancers arising from IBD tissues. MiR-224 levels increased successively at each stage of IBD progression and accurately discriminated cancers from normal or chronically inflamed IBD tissues. Moreover, mRNA arrays combined with bioinformatic analyses suggested the participation of miR-224 in cell cycle regulation. Subsequently, cell cycle experiments indicated that miR-224 regulates the G1-S checkpoint. Finally, in silico prediction analyses, confirmed by Western blotting and luciferase assays, identified p21 as a specific direct mRNA target of miR-224.
  
  
  CONCLUSIONS
  These findings reveal miR dysregulation specific to IBD-associated colorectal carcinoma. MiR-224 is overexpressed in IBD cancers and targets p21, a key cell cycle regulator. Moreover, these results establish the participation of miR-224 in IBD carcinogenesis.
authors: Division of Gastroenterology and Hepatology, Department of Medicine, Johns Hopkins University, Baltimore, Maryland 21231, USA.
authors: Alexandru V
authors: AV
authors: Olaru
authors: Sumitaka
authors: S
authors: Yamanaka
authors: Christine
authors: C
authors: Vazquez
authors: Yuriko
authors: Y
authors: Mori
authors: Yulan
authors: Y
authors: Cheng
authors: John M
authors: JM
authors: Abraham
authors: Theodore M
authors: TM
authors: Bayless
authors: Noam
authors: N
authors: Harpaz
authors: Florin M
authors: FM
authors: Selaru
authors: Stephen J
authors: SJ
authors: Meltzer
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D016022:Case-Control Studies; D015331:Cohort Studies; D003110:Colonic Neoplasms / Q000209:etiology / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism*; D018450:Disease Progression; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D005819:Genetic Markers; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000150:complications* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D035683:MicroRNAs / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012372:ROC Curve; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D015415:Biomarkers; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D005819:Genetic Markers; C546140:MIRN224 microRNA, human; D035683:MicroRNAs
references: Gut. 1994 May;35(5):587-9
references: 8200547
references: Mol Endocrinol. 2010 Mar;24(3):540-51
references: 20118412
references: Gene. 1991 Jun 15;102(1):39-44
references: 1864507
references: PLoS One. 2010 Oct 05;5(10):
references: 20957151
references: PLoS One. 2011 Apr 12;6(4):e18501
references: 21532750
references: BMC Syst Biol. 2010 Apr 27;4:51
references: 20420713
references: Gastroenterology. 2011 Nov;141(5):1749-61.e1
references: 21806946
references: Proc Natl Acad Sci U S A. 2010 Oct 26;107(43):18545-50
references: 20876136
references: Scand J Gastroenterol. 2009;44(1):46-55
references: 18609187
references: J Gastroenterol Hepatol. 2010 May;25 Suppl 1:S129-33
references: 20586854
references: Colorectal Dis. 2011 Nov;13(11):1230-5
references: 21689324
references: Inflamm Bowel Dis. 2011 Jan;17(1):221-31
references: 20848542
references: Gastroenterol Clin North Am. 1989 Mar;18(1):83-97
references: 2646223
references: Nature. 2005 Feb 17;433(7027):769-73
references: 15685193
references: Inflamm Bowel Dis. 2010 Oct;16(10):1729-38
references: 20848482
references: Inflamm Bowel Dis. 2011 Jan;17(1):241-50
references: 20812331
references: Gastroenterology. 2008 Nov;135(5):1624-1635.e24
references: 18835392
references: Gastroenterology. 2003 Nov;125(5):1311-9
references: 14598247
references: Br J Cancer. 2010 Apr 13;102(8):1244-53
references: 20354523
references: N Engl J Med. 1990 Nov 1;323(18):1228-33
references: 2215606
references: Cell. 2001 Dec 28;107(7):823-6
references: 11779458
references: Proc Natl Acad Sci U S A. 2006 Feb 14;103(7):2257-61
references: 16461460
references: Int J Colorectal Dis. 2007 Apr;22(4):411-7
references: 16847674
references: Mol Cell. 2007 Jun 8;26(5):745-52
references: 17540599
references: Aliment Pharmacol Ther. 2003 Sep;18 Suppl 2:1-5
references: 12950413
references: J Clin Endocrinol Metab. 2008 May;93(5):1600-8
references: 18270258
references: Oncology. 2007;72(5-6):397-402
references: 18196926
references: Cancer. 2001 Feb 15;91(4):854-62
references: 11241255
references: Mol Cancer Ther. 2010 Dec;9(12):3105-14
references: 21159603
references: Hepatology. 2011 Dec;54(6):2089-98
references: 21809359
references: Expert Opin Biol Ther. 2011 Feb;11(2):145-55
references: 21219233
references: J Cancer Res Ther. 2010 Oct-Dec;6(4):521-9
references: 21358093
references: J Gastroenterol. 2009;44 Suppl 19:18-22
references: 19148788
references: Cancer Res. 2007 Aug 15;67(16):7713-22
references: 17699775
references: J Biol Chem. 2008 May 9;283(19):13205-15
references: 18319255
references: J Pathol. 2000 Mar;190(4):412-6
references: 10699988
references: Mt Sinai J Med. 2000 May;67(3):227-40
references: 10828908
references: Cancer Genet Cytogenet. 2010 Jul 15;200(2):154-60
references: 20620599
references: Ann Surg Oncol. 2009 Aug;16(8):2339-50
references: 19475450
references: Inflamm Bowel Dis. 2012 Jan;18(1):94-100
references: 21557394
references: PLoS One. 2011;6(11):e27136
references: 22069497
references: Nat Rev Genet. 2008 Feb;9(2):102-14
references: 18197166
references: Hepatology. 2008 Jun;47(6):1955-63
references: 18433021
references: Gut. 2009 Oct;58(10):1375-81
references: 19201770
references: Cancer Sci. 2011 Dec;102(12):2264-71
references: 21883694
references: Cell. 2008 Jan 11;132(1):9-14
references: 18191211
references: Gut. 2012 May;61(5):739-45
references: 21930727