A Prospective Proteomic-Based Study for Identifying Potential Biomarkers for the Diagnosis of Cholangiocarcinoma

BACKGROUND: Cholangiocarcinoma (CCA) is becoming a common fatal hepatic tumor. Early detection of CCA is hampered by the absence of a sufficiently accurate and noninvasive diagnostic test. Proteomic analysis would be a powerful tool to identify potential biomarkers of this cancer. AIMS: This study aims to identify new protein markers that are specific for CCA using proteomic approaches and to evaluate the performance of S100 calcium-binding protein A9 (S100A9) and chaperonin-containing TCR1, subunit 3 (CCTγ) as diagnostic markers for screening test of CCA. METHODS: Two-dimensional differential gel electrophoresis (2-D DIGE) coupled with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry were used to analyze and screen biomarker candidates in the proteomes of five human CCA samples and five healthy control samples. Subsequently, two potential biomarkers, S100A9 and CCTγ, were chosen for validation and analysis by immunohistochemical methods using CCA tissue microarrays. RESULTS: Twenty protein spots were significantly elevated and five protein spots were downregulated in all patients (p < 0.05). The positive rate was significantly higher in patients with CCA (48 ± 35 %) compared with the normal liver control group (5 ± 10 %, p < 0.001), the hepatocellular carcinoma group (15 ± 20 %, p < 0.001), and the cirrhosis group (12 ± 16 %, p < 0.001). A greater proportion of patients with CCA were positive for CCTγ (72 ± 18 %) compared with the normal liver control group (43 ± 22 %, p < 0.001), the hepatocellular carcinoma group (45 ± 20 %, p < 0.001), and the cirrhosis group (39 ± 25 %, p < 0.001). CONCLUSIONS: Combined comparative proteomic analysis using 2-D DIGE and MALDI-TOF is an effective method for identifying differentially expressed proteins in CCA tissues. The expression of S100A9 and CCTγ showed promise as novel diagnostic markers for CCA.