Wisteria floribunda agglutinin-positive mucin 1 is a sensitive biliary marker for human cholangiocarcinoma

Abstract

Cholangiocarcinoma (CC) is an aggressive malignant tumor for which useful markers are not presently available for early and precise diagnosis. The aim of this study was therefore to identify a high-performance diagnostic marker with a special focus on glyco-alteration of glycoproteins. In the course of study, we found that Wisteria floribunda agglutinin (WFA) is the best probe to differentiate intrahepatic cholangiocarcinoma (ICC) lesions from normal bile duct epithelia (BDE) (P < 0.0001). The subsequent histochemical study confirmed ICC-specific WFA staining on 165 tissue specimens. On the other hand, the WFA staining was shown to be closely associated with that of MY.1E12 established previously against sialylated mucin 1 (MUC1) by double-staining experiments. Moreover, glyco-alteration of MUC1 could be verified by western blotting of WFA-captured bile samples from patients with CC patients. Thus, we attempted to construct an enzyme-linked immunosorbent assay system for more convenient CC diagnosis, where WFA-coated plates, the specific monoclonal antibody MY.1E12, and the bile specimens from CC including ICC (n 5 30) and benign diseases (n 5 38) were combined. As a result, CC was clearly distinguished from benign diseases with statistical scores (sensitivity 5 90.0%, specificity 5 76.3%, and area under the curve 5 0.85). As a particular note, the obtained sensitivity is the highest score among those having been so far reported. Conclusion: Our approach focusing significant glyco-alteration of a particular glycoprotein yielded a novel diagnostic system for CC with satisfactory clinical scores. (HEPATOLOGY 2010;52:174-182) C holangiocarcinoma (CC) is an aggressive malignant tumor arising from the epithelial lining of the intrahepatic biliary tract. Although it contributes to only 15% of the total incidence of primary liver cancer, 1 recent epidemiological reports show that the CC incidence has increased significantly in Abbreviations: AFP, alpha-fetoprotein; AUC, area under the curve; BDE, bile duct epithelia; BSA, bovine serum albumin; CA19-9, carbohydrate antigen 19-9;

Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin

“…Thus, its cross-reactive potential severely limits its use for cancer biomarker detection. WFA is currently considered the most prominent diagnostic lectin against cholangiocarcinoma when compared with other homologous lectins (26,35) and the most discriminatory against altered N-glycans on Mac2-binding protein, a secretory N-glycoprotein with elevated expression levels during viral hepatitis induced liver cirrhosis (36,37).…”

Glycome Diagnosis of Human Induced Pluripotent Stem Cells Using Lectin Microarray

“…Analytical protocols using lectin microarray have been developed for various sample types: free oligosaccharides (14,15), tissue sections (16), cell membrane hydrophobic fractions (17,18), and even whole cells (19,20). This technology has just begun to be applied to a wide variety of biological researches, including virus profiling (21) and cell profiling (17,20), and development of cancer glycobiomarkers (22)(23)(24). For cell profiling, less than 100 ng of proteins in hydrophobic fractions are sufficient for each analysis (25,26).…”

Lectin microarray technology identifies specific lectins related to lymph node metastasis of advanced gastric cancer

“…Fluorescently labeled proteins were prepared as previously described [16,17]. Briefly, macrodissected tissue samples (about 5 mm square) were collected in 10 mM citrate buffer (pH 6.0) and incubated for 1 h at 95°C (antigen retrieval).…”