Identification of novel immunohistochemical tumor markers for primary hepatocellular carcinoma; clathrin heavy chain and formiminotransferase cyclodeaminase†

Abstract

Early diagnosis of hepatocellular carcinoma (HCC) greatly improves its prognosis. However, the distinction between benign and malignant tumors is often difficult, and novel immunohistochemical markers are necessary. Using agarose two-dimensional fluorescence difference gel electrophoresis, we analyzed HCC tissues from 10 patients. The fluorescence volumes of 48 spots increased and 79 spots decreased in tumor tissues compared with adjacent nontumor tissue, and 83 proteins were identified by mass spectrometry. Immunoblot confirmed that the expression of clathrin heavy chain (CHC) and Ku86 significantly increased, whereas formiminotransferase cyclodeaminase (FTCD), rhodanese, and vinculin decreased in tumor. The protein expression in tumor and nontumor tissues was further evaluated by immunostaining. Interestingly, CHC and FTCD expression was strikingly different between tumor and nontumor tissues. The sensitivity and specificity of individual markers or a combination for the detection of HCC were 51.8% and 95.6% for CHC, 61.4% and 98.5% for FTCD, and 80.7% and 94.1% for CHC؉FTCD, respectively. Strikingly, the sensitivity and specificity increased to 86.7% and 95.6% when glypican-3, another potential biomarker for HCC, was used with FTCD. Moreover, CHC and FTCD were useful to distinguish early HCC from benign tumors such as regenerative nodule or focal nodular hyperplasia, because the sensitivity and specificity of the markers are 41.2% and 77.8% for CHC, 44.4% and 80.0% for FTCD, which is comparable with those of glypican-3 (33.3% and 100%). The sensitivity significantly increased by combination of these markers, 72.2% for CHC؉FTCD, and 61.1% for CHC؉glypican-3 and FTCD؉glypican-3, as 44.4% of glypican-3 negative early HCC were able to be detected by either CHC or FTCD staining. Conclusion

COP35, a Cholangiocarcinoma-Binding Oligopeptide, Interacts with the Clathrin Heavy Chain Accompanied by GRP78

“…The result supports our hypothesis that COP35 is internalized via clathrin-mediated endocytosis. It was also suggested that CHC is upregulated in hepatocellular carcinoma tissues (30). Consistent with this, our data suggest that CHC expression in CCA lesions was higher than that in normal bile duct epithelium by immunohistochemical staining of formalin-fixed and paraffin-embedded tissues (Table 1; Fig.…”

SAP155-Mediated Splicing of FUSE-Binding Protein-Interacting Repressor Serves as a Molecular Switch for c-myc Gene Expression

“…One was GeLC-MS (18)(19)(20) ). The minimum criteria for protein identification were protein and peptide thresholds set to 95.0% (Scaffold's probability threshold filter) and number of unique peptides set to 2.…”

The application of markers (HSP70 GPC3 and GS) in liver biopsies is useful for detection of hepatocellular carcinoma

“…Unfortunately genes which prove useful in expression studies not always can be translated in robust tissue markers and used in the routine practice as immunocytochemical tools. We believe our panel susceptible to be improved by the addition of novel markers [11,23] as to further increase the reliability and objectivity of HCC diagnosis in liver biopsies and cytological specimens and to meet the growing expectancy of liver clinicians, surgeons and oncologists.…”