title: iTRAQ™ labeling coupled with LC-MALDI mass spectrometry for monitoring temporal response of colorectal cancer cells to butyrate treatment.
abstract: Mass spectrometry (MS)-based quantitative proteomics plays important roles in drug discovery. In this chapter, we describe a stable isotope labeling technique which employs 4-plex iTRAQ(™) isobaric reagents coupled with two-dimensional (2-D) liquid chromatography (LC) and MALDI-TOF/TOF MS, for a temporal study of HCT-116 colon carcinoma cells treated with butyrate. Butyrate is a short-chain fatty acid fermentation by-product of fiber that can induce temporal cell maturation, from the early phase of growth arrest, to differentiation, and to the activation of apoptotic cascades. Our quantitative proteomics study uncovered several integrated cellular processes and pathways involved in growth arrest, apoptosis, and metastasis. Selected protein targets are validated by real-time PCR and western blotting.
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
authors.forename: Hwee Tong
authors.initials: HT
authors.lastname: Tan
authors.forename: Teck Kwang
authors.initials: TK
authors.lastname: Lim
authors.forename: Maxey C M
authors.initials: MC
authors.lastname: Chung
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Lin
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D015153:Blotting, Western / Q000379:methods; D002087:Butyrates / Q000494:pharmacology* / Q000627:therapeutic use; D002277:Carcinoma / Q000188:drug therapy*; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods; D015179:Colorectal Neoplasms / Q000188:drug therapy*; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D020543:Proteome / Q000187:drug effects* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D000970:Antineoplastic Agents; D002087:Butyrates; D020543:Proteome