title: CD109 Overexpression in Pancreatic Cancer Identified by Cell-Surface Glycoprotein Capture.
abstract: BACKGROUND
  The development of novel targeted cancer therapies and/or diagnostic tools is dependent upon an understanding of the differential expression of molecular targets between normal tissues and tumors. Many of these potential targets are cell-surface receptors; however, our knowledge of the cell-surface proteins upregulated in pancreatic tumors is limited, thus impeding the development of targeted therapies for pancreatic cancer. To develop new diagnostic and therapeutic tools to specifically target pancreatic tumors, we sought to identify cell-surface proteins that may serve as potential tumor-specfic targets.
  
  
  METHODS
  Membrane glycoproteins on the pancreatic cancer cell lines BxPC-3 were labeled with the bifunctional linker biocytin hydrazide. Following proteolytic digestion, biotinylated glycopeptides were captured with streptavidin-coupled beads then released by PNGaseF-mediated endoglycosidase cleavage and identified by liquid chromatography-tandem mass spectrometry (MS). A protein identified by the cell-surface glycoprotein capture procedure, CD109, was evaluated by western analysis of lysates of pancreatic cancer cell lines and by immunohistochemistry in sections of pancreatic ductal adenocarcinoma and non- neoplastic pancreatic tissues.
  
  
  RESULTS
  MS/MS analysis of glycopeptides captured from BxPC-3 cells revealed 18 proteins predicted or known to be associated with the plasma membrane, including CD109, which has not been reported in pancreatic cancer. Western analysis of CD109 in lysates prepared from pancreatic cancer cell lines revealed it was expressed in 6 of 8 cell lines, with a high level of expression in BxPC-3, MIAPaCa-2, and Panc-1 cells. Immunohistochemical analyses of human pancreatic tissues indicate CD109 is significantly overexpressed in pancreatic tumors compared to normal pancreas.
  
  
  CONCLUSIONS
  The selective capture of glycopeptides from the surface of pancreatic cancer cell lines can reveal novel cell-surface glycoproteins expressed in pancreatic ductal adenocarcinomas.
authors: Central Arkansas Veterans Healthcare System, University of Arkansas for Medical Sciences; Little Rock, Arkansas, USA ; Departments of Pharmaceutical Sciences, University of Arkansas for Medical Sciences; Little Rock, Arkansas, USA.
authors: Randy S
authors: RS
authors: Haun
authors: Central Arkansas Veterans Healthcare System, University of Arkansas for Medical Sciences; Little Rock, Arkansas, USA.
authors: Chun-Yang
authors: CY
authors: Fan
authors: Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences; Little Rock, Arkansas, USA.
authors: Samuel G
authors: SG
authors: Mackintosh
authors: Department of Experimental Center of Functional Subjects, College of Basic Medicine, China Medical University, Shenyang, China.
authors: Hong
authors: H
authors: Zhao
authors: Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences; Little Rock, Arkansas, USA.
authors: Alan J
authors: AJ
authors: Tackett
keywords: Glycoproteins; Pancreatic cancer; Proteomic profiling
references: Am J Pathol. 2003 Apr;162(4):1151-62
references: 12651607
references: Pathol Int. 2008 May;58(5):288-94
references: 18429827
references: J Exp Clin Cancer Res. 2012 Aug 27;31:68
references: 22925330
references: World J Gastroenterol. 2003 Apr;9(4):818-23
references: 12679940
references: Appl Biochem Biotechnol. 2012 Aug;167(8):2257-68
references: 22695923
references: Nat Biotechnol. 2009 Apr;27(4):378-86
references: 19349973
references: Oncogene. 2004 Apr 29;23(20):3716-20
references: 15116102
references: Mol Cell Proteomics. 2009 Nov;8(11):2555-69
references: 19656770
references: Biochem Biophys Res Commun. 2002 Apr 26;293(1):391-5
references: 12054612
references: Mol Cell Proteomics. 2006 Jan;5(1):157-71
references: 16215274
references: Cancer Res. 2014 Jun 1;74(11):2913-21
references: 24840647
references: Cancer Res. 2005 Mar 1;65(5):1619-26
references: 15753353
references: Cancer Invest. 1986;4(1):15-23
references: 3754176
references: Clin Cancer Res. 2005 Apr 15;11(8):2840-52
references: 15837731
references: Mol Carcinog. 2006 Nov;45(11):814-27
references: 16865675
references: Aliment Pharmacol Ther. 2003 Dec;18(11-12):1049-69
references: 14653825
references: Int J Cancer. 2004 Oct 20;112(1):100-12
references: 15305381
references: Nat Rev Gastroenterol Hepatol. 2011 Dec 20;9(2):77-87
references: 22183185
references: Neoplasia. 2004 Sep-Oct;6(5):611-22
references: 15548371
references: Oncogene. 2002 Jul 4;21(29):4587-94
references: 12085237
references: Mol Cell Proteomics. 2007 Aug;6(8):1331-42
references: 17496331
references: Am J Pathol. 2002 Jan;160(1):91-9
references: 11786403
references: Pancreatology. 2004;4(6):587-97
references: 15557762
references: Cancer Res. 2002 May 15;62(10):2890-6
references: 12019169
references: Blood. 2002 Mar 1;99(5):1683-91
references: 11861284
references: Jpn J Cancer Res. 1987 Jan;78(1):54-62
references: 3102439
references: J Pathol. 2003 Sep;201(1):63-74
references: 12950018
references: Cancer Res. 2003 May 15;63(10):2649-57
references: 12750293
references: Oncogene. 2004 Mar 25;23(13):2385-400
references: 14767473
references: Int J Colorectal Dis. 2004 Sep;19(5):401-13
references: 14745573
references: Mol Cell Proteomics. 2009 Apr;8(4):624-38
references: 19036722
references: CA Cancer J Clin. 2014 Jan-Feb;64(1):9-29
references: 24399786
references: Mol Cell Biol. 1999 Mar;19(3):1720-30
references: 10022859
references: Cell Mol Life Sci. 2003 Jun;60(6):1180-99
references: 12861384
references: Gastroenterology. 2005 Nov;129(5):1454-63
references: 16285947
references: Cancer Res. 2003 Dec 15;63(24):8614-22
references: 14695172
references: Am J Pathol. 2002 Apr;160(4):1239-49
references: 11943709