Proteins Secreted by Lung Adenocarcinoma Cells -Identification of Candidate Diagnosis Markers from Culture Supernatants of Dissected Tumor Tissue-

目的.肺癌の早期診断に有用なバイオマーカーの確立は治療成績向上のための重要な課題である.我々は血清マーカーを探索する目的で,肺腺癌組織から分泌あるいは漏出する一群の蛋白質を分析した.方法.外科的に切除した12症例の肺腺癌組織および同じ症例の正常な末梢肺組織を各々無血清培地で短期培養した.培養上清中の癌組織および正常組織由来の蛋白質を,異なる蛍光波長を持つ色素化合物(CyDyeTM:Cy2,Cy3 and Cy5)でそれぞれ標識した.標識試料を症例ごとに混合後,各々二次元電気泳動で展開した.結果.二次元ゲル上で検出された蛋白質スポットのうち,34個のスポットは少なくとも5症例から検出され,かつ正常組織に対する癌組織の平均蛍光強度比が2.0以上を示した(p<0.05).質量分析を含む解析手順により34スポットのすべてから有意な蛋白質同定結果を得た.同定された蛋白質には,napsin Aやpulmonary surfactant-associated protein Aなど,II型肺胞上皮細胞への組織学的分化を示す蛋白質が含まれていた.また,腫瘍組織全般に共通して発現の亢進することが示唆される蛋白質も同定された.結論.組織培養からのアプローチは肺腺癌の血清マーカーの候補蛋白質を同定するために有効である.

[1]  E. Diamandis,et al.  Identification of Five Candidate Lung Cancer Biomarkers by Proteomics Analysis of Conditioned Media of Four Lung Cancer Cell Lines* , 2009, Molecular & Cellular Proteomics.

[2]  E. Diamandis,et al.  Comprehensive analysis of conditioned media from ovarian cancer cell lines identifies novel candidate markers of epithelial ovarian cancer. , 2009, Journal of proteome research.

[3]  A. Jemal,et al.  Cancer Statistics, 2009 , 2009, CA: a cancer journal for clinicians.

[4]  Shinji Sato,et al.  Amino Acid Sequence Database Suitable for the Protein and Proteome Analysis , 2008 .

[5]  M. Páez de la Cadena,et al.  How the measurements of a few serum markers can be combined to enhance their clinical values in the management of cancer. , 2008, Anticancer research.

[6]  H. Iso,et al.  Sensitivity and specificity of lung cancer screening using chest low-dose computed tomography , 2008, British Journal of Cancer.

[7]  H. Antti,et al.  Evaluation of 2‐D DIGE for skeletal muscle: Protocol and repeatability , 2008, Scandinavian journal of clinical and laboratory investigation.

[8]  David E. Misek,et al.  Identification of 14-3-3θ as an antigen that induces a humoral response in lung cancer , 2007 .

[9]  Alissa K. Greenberg,et al.  Biomarkers for lung cancer: clinical uses , 2007, Current opinion in pulmonary medicine.

[10]  A. Jemal,et al.  Reduced lung cancer deaths attributable to decreased tobacco use in Massachusetts , 2007, Cancer Causes & Control.

[11]  J. Jett Limitations of Screening for Lung Cancer with Low-Dose Spiral Computed Tomography , 2005, Clinical Cancer Research.

[12]  H. Kato,et al.  Clinical‐scale high‐throughput human plasma proteome analysis: Lung adenocarcinoma , 2005, Proteomics.

[13]  Toshihide Nishimura,et al.  Protein identification from product ion spectra of peptides validated by correlation between measured and predicted elution times in liquid chromatography/mass spectrometry , 2005, Proteomics.

[14]  N. Anderson,et al.  The Human Plasma Proteome: History, Character, and Diagnostic Prospects , 2003, Molecular & Cellular Proteomics.

[15]  N. Anderson,et al.  The Human Plasma Proteome , 2002, Molecular & Cellular Proteomics.

[16]  M. Senko,et al.  A two-dimensional quadrupole ion trap mass spectrometer , 2002, Journal of the American Society for Mass Spectrometry.

[17]  W. Peters,et al.  PLASMA GLUTATHIONE S-TRANSFERASE π1-1 AND α1-1 LEVELS IN PATIENTS WITH BLADDER CANCER , 2000 .

[18]  H. Kato,et al.  Human Tissue Distribution of TA02, Which is Homologous with a New Type of Aspartic Proteinase, Napsin A , 2000, Japanese journal of cancer research : Gann.

[19]  D. N. Perkins,et al.  Probability‐based protein identification by searching sequence databases using mass spectrometry data , 1999, Electrophoresis.

[20]  A Charloux,et al.  The increasing incidence of lung adenocarcinoma: reality or artefact? A review of the epidemiology of lung adenocarcinoma. , 1997, International journal of epidemiology.

[21]  G. Paludetti,et al.  Cathepsin D concentration in primary laryngeal cancer: Correlation with clinico‐pathological parameters, EGFR status and prognosis , 1996, International journal of cancer.

[22]  A. Shevchenko,et al.  Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. , 1996, Analytical chemistry.

[23]  Tohru Takahashi,et al.  Atypical adenomatous hyperplasia and adenocarcinoma of the human lung: Their heterology in form and analogy in immunohistochemical characteristics , 1996, Cancer.

[24]  N. Kohno,et al.  Detection of soluble tumor-associated antigens in sera and effusions using novel monoclonal antibodies, KL-3 and KL-6, against lung adenocarcinoma. , 1988, Japanese journal of clinical oncology.

[25]  David E. Misek,et al.  Identification of 14-3-3 theta as an antigen that induces a humoral response in lung cancer. , 2007, Cancer research.

[26]  W. Peters,et al.  Plasma glutathione S-transferase pi 1-1 AND alpha 1-1 levels in patients with bladder cancer. , 2000, The Journal of urology.

[27]  M. Ünlü,et al.  Difference gel electrophoresis. A single gel method for detecting changes in protein extracts , 1997, Electrophoresis.

[28]  D. Hochstrasser,et al.  Improved and simplified in‐gel sample application using reswelling of dry immobilized pH gradients , 1997, Electrophoresis.