title: Quantitative LC-MS/MS Analysis of Proteins Involved in Metastasis of Breast Cancer.
abstract: The purpose of this study was to develop quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for the analysis of proteins involved in metastasis of breast cancer for diagnosis and determining disease prognosis, as well as to further our understand of metastatic mechanisms. We have previously demonstrated that the protein type XIV collagen may be specifically expressed in metastatic tissues by two dimensional LC-MS/MS. In this study, we developed quantitative LC-MS/MS methods for type XIV collagen. Type XIV collagen was quantified by analyzing 2 peptides generated by digesting type XIV collagen using stable isotope-labeled peptides. The individual concentrations were equivalent between 2 different peptides of type XIV collagen by evaluation of imprecise transitions and using the best transition for the peptide concentration. The results indicated that type XIV collagen is highly expressed in metastatic tissues of patients with massive lymph node involvement compared to non-metastatic tissues. These findings were validated by quantitative real-time RT-PCR. Further studies on type XIV collagen are desired to verify its role as a prognostic factor and diagnosis marker for metastasis.
authors.affiliation: Department of Clinical Laboratory Medicine, Wakayama Medical University,Wakayama, Japan; JCL Bioassay Corporation, Nishiwaki, Hyogo, Japan.
authors.forename: Rieko
authors.initials: R
authors.lastname: Goto
authors.affiliation: Department of Clinical Laboratory Medicine, Wakayama Medical University,Wakayama, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Nakamura
authors.affiliation: JCL Bioassay Corporation, Nishiwaki, Hyogo, Japan.
authors.forename: Tomonori
authors.initials: T
authors.lastname: Takami
authors.affiliation: Department of Clinical Laboratory Medicine, Wakayama Medical University,Wakayama, Japan.
authors.forename: Tokio
authors.initials: T
authors.lastname: Sanke
authors.affiliation: Graduate School of Pharmaceutical Science Osaka University, Suita, Osaka, Japan.
authors.forename: Zenzaburo
authors.initials: Z
authors.lastname: Tozuka
mesh_terms: D000595:Amino Acid Sequence; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D002853:Chromatography, Liquid / Q000379:methods* / Q000592:standards; D003094:Collagen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008207:Lymphatic Metastasis; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011379:Prognosis; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards
chemical_list: C054447:COL14A1 protein, human; D006023:Glycoproteins; D009363:Neoplasm Proteins; D010455:Peptides; D003094:Collagen