title: Biomarker discovery from pancreatic cancer secretome using a differential proteomic approach.
abstract: Quantitative proteomics can be used as a screening tool for identification of differentially expressed proteins as potential biomarkers for cancers. Candidate biomarkers from such studies can subsequently be tested using other techniques for use in early detection of cancers. Here we demonstrate the use of stable isotope labeling with amino acids in cell culture (SILAC) method to compare the secreted proteins (secretome) from pancreatic cancer-derived cells with that from non-neoplastic pancreatic ductal cells. We identified 145 differentially secreted proteins (>1.5-fold change), several of which were previously reported as either up-regulated (e.g. cathepsin D, macrophage colony stimulation factor, and fibronectin receptor) or down-regulated (e.g. profilin 1 and IGFBP-7) proteins in pancreatic cancer, confirming the validity of our approach. In addition, we identified several proteins that have not been correlated previously with pancreatic cancer including perlecan (HSPG2), CD9 antigen, fibronectin receptor (integrin beta1), and a novel cytokine designated as predicted osteoblast protein (FAM3C). The differential expression of a subset of these novel proteins was validated by Western blot analysis. In addition, overexpression of several proteins not described previously to be elevated in human pancreatic cancer (CD9, perlecan, SDF4, apoE, and fibronectin receptor) was confirmed by immunohistochemical labeling using pancreatic cancer tissue microarrays suggesting that these could be further pursued as potential biomarkers. Lastly the protein expression data from SILAC were compared with mRNA expression data obtained using gene expression microarrays for the two cell lines (Panc1 and human pancreatic duct epithelial), and a correlation coefficient (r) of 0.28 was obtained, confirming previously reported poor associations between RNA and protein expression studies.
authors.affiliation: Department of Biological Chemistry, McKusick-Nathans Institute of Genetic Medicine, The Johns Hopkins University, Baltimore, Maryland 21205, USA.
authors.forename: Mads
authors.initials: M
authors.lastname: Grønborg
authors.forename: Troels Zakarias
authors.initials: TZ
authors.lastname: Kristiansen
authors.forename: Akiko
authors.initials: A
authors.lastname: Iwahori
authors.forename: Rubens
authors.initials: R
authors.lastname: Chang
authors.forename: Raghunath
authors.initials: R
authors.lastname: Reddy
authors.forename: Norihiro
authors.initials: N
authors.lastname: Sato
authors.forename: Henrik
authors.initials: H
authors.lastname: Molina
authors.forename: Ole Nørregaard
authors.initials: ON
authors.lastname: Jensen
authors.forename: Ralph H
authors.initials: RH
authors.lastname: Hruban
authors.forename: Michael G
authors.initials: MG
authors.lastname: Goggins
authors.forename: Anirban
authors.initials: A
authors.lastname: Maitra
authors.forename: Akhilesh
authors.initials: A
authors.lastname: Pandey
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism*; D015153:Blotting, Western; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D004847:Epithelial Cells / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D007553:Isotope Labeling; D020411:Oligonucleotide Array Sequence Analysis; D010183:Pancreatic Ducts / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism*; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D014408:Biomarkers, Tumor; D020543:Proteome