title: Phosphoproteomics of colon cancer metastasis: comparative mass spectrometric analysis of the isogenic primary and metastatic cell lines SW480 and SW620.
abstract: The contributions of phosphorylation-mediated signaling networks to colon cancer metastasis are poorly defined. To interrogate constitutive signaling alterations in cancer progression, the global phosphoproteomes of patient-matched SW480 (primary colon tumor origin) and SW620 (lymph node metastasis) cell lines were compared with TiO2and immobilized metal affinity chromatography phosphopeptide enrichment followed by liquid chromatography-tandem mass spectrometry. Network analysis of the significantly altered phosphosites revealed differential regulation in cellular adhesion, mitosis, and messenger RNA translational machinery. Messenger RNA biogenesis and splicing, transport through the nuclear pores, initiation of translation, and stability and degradation were also affected. Although alterations in these processes have been associated with oncogenic transformation, control of messenger RNA stability has typically not been associated with cancer progression. Notably, the single phosphosite with the greatest relative change in SW620 cells was Ser2 on eukaryotic translation initiation factor 2 subunit 2, suggesting that SW620 cells translate faster or with greater efficiency than SW480 cells. These broad changes in the regulation of translation also occur without overexpression of eukaryotic translation initiation factor 4E. The findings suggest that metastatic cells exhibit constitutive changes to the phosphoproteome, and that messenger RNA stability and translational efficiency may be important targets of deregulation during cancer progression.
authors.affiliation: Department of Chemistry and Biochemistry, Harper Cancer Research Institute, University of Notre Dame, 251 Nieuwland Science Hall, Notre Dame, IN, 46556, USA.
authors.forename: Alissa J
authors.initials: AJ
authors.lastname: Schunter
authors.affiliation: Department of Chemistry and Biochemistry, Harper Cancer Research Institute, University of Notre Dame, 251 Nieuwland Science Hall, Notre Dame, IN, 46556, USA.
authors.forename: Xiaoshan
authors.initials: X
authors.lastname: Yue
authors.affiliation: Department of Chemistry and Biochemistry, Harper Cancer Research Institute, University of Notre Dame, 251 Nieuwland Science Hall, Notre Dame, IN, 46556, USA. ahummon@nd.edu.
authors.forename: Amanda B
authors.initials: AB
authors.lastname: Hummon
mesh_terms: D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000473:pathology*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009362:Neoplasm Metastasis; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D014176:Protein Biosynthesis; D040901:Proteomics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D010750:Phosphoproteins; D012333:RNA, Messenger
keywords: Colon cancer progression; Immobilized metal affinity chromatography; Immunoprecipitation–selected reaction monitoring; Messenger RNA translation; Phosphosite stoichiometry; TiO2