Proteomic analysis of laser-microdissected paraffin-embedded tissues: (1) Stage-related protein candidates upon non-metastatic lung adenocarcinoma

• Published in: Journal of proteomics 2010-04, Vol.73 (6), p.1089-1099
• Main Authors: Kawamura, Takeshi, Nomura, Masaharu, Tojo, Hiromasa, Fujii, Kiyonaga, Hamasaki, Hiroko, Mikami, Sayaka, Bando, Yasuhiko, Kato, Harubumi, Nishimura, Toshihide
• Format: Article
• Language: English
• Subjects: Adenocarcinoma; Biological and medical sciences; biomarkers; Cell physiology; Cell transformation and carcinogenesis. Action of oncogenes and antioncogenes; Diverse techniques; Enumeration; Formalin-fixed paraffin-embedded tissue; Fundamental and applied biological sciences. Psychology; Laser microdissection; Lasers; Liquid chromatography; Lung adenocarcinoma; Lung cancer; Lymph nodes; Mass spectrometry; Mass spectroscopy; Medical sciences; Molecular and cellular biology; Pneumology; proteomics; Quantitation; Solubilization; Spectral counting; Stage-related biomarkers; Statistical analysis; Trypsin; Tumors of the respiratory system and mediastinum
• ISSN: 1874-3919
• DOI: 10.1016/j.jprot.2009.11.011

We used formalin-fixed paraffin-embedded (FFPE) materials for biomarker discovery in cases of lung cancer using proteomic analysis. We conducted a retrospective global proteomic study in order to characterize protein expression reflecting clinical stages of individual patients with stage I lung adenocarcinoma without lymph node involvement ( n = 7). In addition, we studied more advanced stage IIIA with spread to lymph nodes ( n = 6), because the degree of lymph node involvement is the most important factor for staging. FFPE sections of cancerous lesions resected surgically from patients with well-characterized clinical history were subjected to laser microdissection (LMD) followed by Liquid Tissue™ solubilization and digestion trypsin. Spectral counting was used to measure the amounts of proteins identified by shotgun liquid chromatography (LC)/tandem mass spectrometry (MS/MS). More than 500 proteins were identified from IA and IIIA cases, and non-parametric statistics showed that 81 proteins correlated significantly with stage IA or IIIA. A subset of those proteins were verified by multiple-reaction monitoring mass spectrometric quantitation (MRM assay), described in other paper in this issue. These results demonstrated the technical feasibility of a global proteomic study using clinically well documented FFPE sections, and its possible utility for detailed retrospective disease analyses in order to improve therapeutic strategy.