title: Identification of key players for colorectal cancer metastasis by iTRAQ quantitative proteomics profiling of isogenic SW480 and SW620 cell lines.
abstract: This study compared the whole cell proteome profiles of two isogenic colorectal cancer (CRC) cell lines (primary SW480 cell line and its lymph node metastatic variant SW620), as an in vitro metastatic model, to gain an insight into the molecular events of CRC metastasis. Using iTRAQ (isobaric tags for relative and absolute quantitation) based shotgun proteomics approach, we identified 1140 unique proteins, out of which 147 were found to be significantly altered in the metastatic cell. Ingenuity pathway analysis with those significantly altered proteins, revealed cellular organization and assembly as the top-ranked altered biological function. Differential expression pattern of 6 candidate proteins were validated by Western blot. Among these, the low expression level of β-catenin combined with the up-regulation of CacyBP (Calcyclin binding Protein), a β-catenin degrading protein, in the metastatic cell provided a rational guide for the downstream functional assays. The relative expression pattern of these two proteins was further validated in three other CRC cells by Western blot and quantitative immunofluorescence studies. Overexpression of CacyBP in three different primary CRC cell lines showed significant reduction in adhesion characteristics as well as cellular β-catenin level as confirmed by our experiments, indicating the possible involvement of CacyBP in CRC metastasis. In short, this study demonstrates successful application of a quantitative proteomics approach to identify novel key players for CRC metastasis, which may serve as biomarkers and/or drug targets to improve CRC therapy.
authors.affiliation: Department of Biological Sciences, National University of Singapore , 14 Science Drive 4, Singapore 117543.
authors.forename: Dipanjana
authors.initials: D
authors.lastname: Ghosh
authors.forename: Han
authors.initials: H
authors.lastname: Yu
authors.forename: Xing Fei
authors.initials: XF
authors.lastname: Tan
authors.forename: Teck Kwang
authors.initials: TK
authors.lastname: Lim
authors.forename: Ramdzan M
authors.initials: RM
authors.lastname: Zubaidah
authors.forename: Hwee Tong
authors.initials: HT
authors.lastname: Tan
authors.forename: Maxey C M
authors.initials: MC
authors.lastname: Chung
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Lin
mesh_terms: D000465:Algorithms; D015415:Biomarkers; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000473:pathology*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008207:Lymphatic Metastasis; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods; D008875:Middle Aged; D009362:Neoplasm Metastasis; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods; D051176:beta Catenin / Q000378:metabolism
chemical_list: D015415:Biomarkers; D051176:beta Catenin