title: Proteomic differences between hepatocellular carcinoma and nontumorous liver tissue investigated by a combined gel-based and label-free quantitative proteomics study.
abstract: Proteomics-based clinical studies have been shown to be promising strategies for the discovery of novel biomarkers of a particular disease. Here, we present a study of hepatocellular carcinoma (HCC) that combines complementary two-dimensional difference in gel electrophoresis (2D-DIGE) and liquid chromatography (LC-MS)-based approaches of quantitative proteomics. In our proteomic experiments, we analyzed a set of 14 samples (7 × HCC versus 7 × nontumorous liver tissue) with both techniques. Thereby we identified 573 proteins that were differentially expressed between the experimental groups. Among these, only 51 differentially expressed proteins were identified irrespective of the applied approach. Using Western blotting and immunohistochemical analysis the regulation patterns of six selected proteins from the study overlap (inorganic pyrophosphatase 1 (PPA1), tumor necrosis factor type 1 receptor-associated protein 1 (TRAP1), betaine-homocysteine S-methyltransferase 1 (BHMT)) were successfully verified within the same sample set. In addition, the up-regulations of selected proteins from the complements of both approaches (major vault protein (MVP), gelsolin (GSN), chloride intracellular channel protein 1 (CLIC1)) were also reproducible. Within a second independent verification set (n = 33) the altered protein expression levels of major vault protein and betaine-homocysteine S-methyltransferase were further confirmed by Western blots quantitatively analyzed via densitometry. For the other candidates slight but nonsignificant trends were detectable in this independent cohort. Based on these results we assume that major vault protein and betaine-homocysteine S-methyltransferase have the potential to act as diagnostic HCC biomarker candidates that are worth to be followed in further validation studies.
authors: Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany.
authors: Dominik A
authors: DA
authors: Megger
authors: Thilo
authors: T
authors: Bracht
authors: Michael
authors: M
authors: Kohl
authors: Maike
authors: M
authors: Ahrens
authors: Wael
authors: W
authors: Naboulsi
authors: Frank
authors: F
authors: Weber
authors: Andreas-Claudius
authors: AC
authors: Hoffmann
authors: Christian
authors: C
authors: Stephan
authors: Katja
authors: K
authors: Kuhlmann
authors: Martin
authors: M
authors: Eisenacher
authors: Jörg F
authors: JF
authors: Schlaak
authors: Hideo A
authors: HA
authors: Baba
authors: Helmut E
authors: HE
authors: Meyer
authors: Barbara
authors: B
authors: Sitek
mesh_terms: D000328:Adult; D000368:Aged; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D008099:Liver / Q000378:metabolism*; D008113:Liver Neoplasms / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D057825:Two-Dimensional Difference Gel Electrophoresis; D055815:Young Adult
chemical_list: D009363:Neoplasm Proteins