Identification of Specific Protein Markers in Microdissected Hepatocellular Carcinoma Christian Melle,† Gu1 nther Ernst,† Olaf Scheibner,‡ Roland Kaufmann,§ Bettina Schimmel,† Annett Bleul,† Utz Settmacher,§ Merten Hommann,§ Uwe Claussen,† and Ferdinand von Eggeling*,† Core Unit Chip Application (CUCA), Institute of Human Genetics and Anthropology, Medical Faculty at the Friedrich Schiller University, 07740 Jena, Germany, Leibniz Institute for Natural Products Research and Infection Biology, Hans-Kno¨ll-Institute, 07745 Jena, Germany, University Hospital, Department of General and Visceral Surgery, Medical Faculty at the Friedrich Schiller University, 07740 Jena, Germany Received August 29, 2006
At present, the molecular mechanisms of hepatocellular carcinogenesis are not well-understood, and hepatocellular carcinoma (HCC) stays one of the most frequent and high-risk metastatic visceral neoplasms worldwide. For the identification of tumor-relevant proteins, we analyzed microdissected cells from nontumorous liver tissue (n ) 28) and tissue derived from hepatic tumor center (n ) 25), as well as tumor margin (n ) 23). We unequivocally identified 53 proteins from hepatic tumor tissues by peptide fingerprint mapping and SELDI mass spectrometry that were separated using two-dimensional gel electrophoresis. Among a number of signals that were detected as significantly different in the protein profiling analysis, we identified for the first time ferritin light subunit (FLS) and adenylate kinase 3 alpha-like 1 (AK3), showing decreased expressions in hepatic tumor, as well as biliverdin reductase B (BVRB) that was upregulated in HCC. The use of ProteinChip technology in combination with tissue microdissection gives insight of the complex changes occurring at the protein level in hepatocellular cancer associated with tumor development and progression and resulted in three new potential diagnostically useful markers. Keywords: hepatocellular carcinoma • proteomics • biomarker • tissue microdissection • SELDI-TOF MS • tandem MS • two-dimensional gel electrophoresis • immunohistochemistry
Introduction Hepatocellular carcinoma (HCC) is one of the most frequent and high-risk metastatic visceral neoplasms worldwide.1 The incidence ranges from
