title: Proteomic analysis of laser-microdissected paraffin-embedded tissues: (2) MRM assay for stage-related proteins upon non-metastatic lung adenocarcinoma.
abstract: A preceding paper suggested 81 candidates of stage-specifically expressed proteins for either stage IA or IIIA by global shotgun proteomics and spectral counting. Six proteins, a subset of these proteins, were chosen for a further verification study since they are potentially soluble and/or secretory, which nature is convenient for detecting them in blood in clinical practice. The multiple-reaction monitoring (MRM) quantitative analysis suggested that napsin-A and anterior gradient protein 2 homolog (hAG-2) out of the 6 candidates would be useful for determining stage IA or IIIA and are related to metastasis. In the study we noted that stage IIIA patients with better outcome showed napsin-A profiles similar to that of stage IA patients. We therefore examined 14 additional patients for analysis, which contained the IA-stage patients of poorer outcome and the IIIA-stage patients of better outcome. The MRM analysis of napsin-A for all patients suggests that napsin-A contents correlate with better outcome in stage IA. This and discovery studies demonstrate that direct isolation of tumor cells alone by laser microdissection (LMD) greatly reduces complexity on comprehensive analyses, and that MRM mass spectrometry using the endogenous internal standard is a feasible technology for quantitative verification of target proteins in formalin-fixed paraffin embedded (FFPE) tissues.
authors: Department of Surgery, Tokyo Medical University, Shinjuku, Tokyo, Japan. linne300@aol.com
authors: Toshihide
authors: T
authors: Nishimura
authors: Masaharu
authors: M
authors: Nomura
authors: Hiromasa
authors: H
authors: Tojo
authors: Hiroko
authors: H
authors: Hamasaki
authors: Tetsuya
authors: T
authors: Fukuda
authors: Kiyonaga
authors: K
authors: Fujii
authors: Sayaka
authors: S
authors: Mikami
authors: Yasuhiko
authors: Y
authors: Bando
authors: Harubumi
authors: H
authors: Kato
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis* / Q000473:pathology; D000328:Adult; D000368:Aged; D016282:Aspartic Acid Endopeptidases / Q000096:biosynthesis; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000473:pathology; D008297:Male; D042282:Microdissection; D008875:Middle Aged; D016612:Paraffin Embedding / Q000379:methods; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D016896:Treatment Outcome
chemical_list: D020543:Proteome; D016282:Aspartic Acid Endopeptidases; C498549:NAPSA protein, human