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N ‐glycoprotein profiling of lung adenocarcinoma pleural effusions by shotgun proteomics
Author(s) -
Soltermann Alex,
Ossola Reto,
KilgusHawelski Sandra,
von Eckardstein Arnold,
Suter Tobias,
Aebersold Ruedi,
Moch Holger
Publication year - 2008
Publication title -
cancer cytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/cncr.23349
Subject(s) - medicine , pleural effusion , lung cancer , adenocarcinoma , periostin , metastasis , pathology , proteomics , biomarker , malignant pleural effusion , effusion , lung , cancer , biology , biochemistry , surgery , gene , extracellular matrix
BACKGROUND Malignant pleural effusion of advanced lung adenocarcinoma may be a valid source for detection of biomarkers, such as N ‐glycosylated proteins (N‐GP), because tumor cells grow during weeks in this liquid. The authors aimed for creation of N‐GP effusion profiles from routine cytology specimens to detect relevant biomarkers. METHODS Hundred microliters of malignant pleural effusions of 5 patients with lung adenocarcinoma and 5 nonmalignant controls were used for triplicate N‐GP capture by solid‐phase extraction. After trypsin digest and PNGase F release, a liquid chromatography separation connected online to a tandem mass spectrometer was performed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). RESULTS In the total of 10 samples, 170 and 278 nonredundant proteins were detected with probabilities of ≥.9 and ≥.5, respectively. The specificity for the N‐glycomotif was 88% at P ≥ .9. Penetration into the moderate to low protein concentration range (μg‐ng/mL) occurred, and several proteins associated with tumor progression or metastasis were identified, including CA‐125, CD44, CD166, lysosome‐associated membrane glycoprotein 2 (LAMP‐2), multimerin 2, and periostin. MS identifications were correlated with the corresponding immunoreactivity in either effusion fluid or tumor tissue. CONCLUSIONS In conclusion, reduction of sample complexity by N‐GP capturing allows detection of proteins in the μg to ng/mL range. Pleural effusion is a useful source for biomarker research in lung cancer. Cancer (Cancer Cytopathol) 2008. © 2008 American Cancer Society.