title: Development of a highly sensitive glycan microarray for quantifying AFP-L3 for early prediction of hepatitis B virus-related hepatocellular carcinoma.
abstract: The α-fetoprotein fraction L3 (AFP-L3), which is synthesized by malignant cells and incorporates a fucosylated oligosaccharide, has been investigated as a diagnostic and prognostic marker for hepatocellular carcinoma (HCC). Quantification of AFP-L3 by conventional enzyme-linked immunosorbent assay (ELISA) has not always produced reliable results for serum samples with low AFP, and thus we evaluated the clinical utility of quantifying AFP-L3 using a new and highly sensitive glycan microarray assay. Sera from 9 patients with chronic hepatitis B and 32 patients with hepatitis B virus (HBV)-related HCC were tested for AFP-L3 level using the glycan microarray. Additionally, we compared receiver operator characteristic curves for the ELISA and glycan microarray methods for determination of the AFP-L3: AFP-L1 ratio in patient samples. This ratio was calculated for 8 HCC patients who underwent transarterial embolization therapy pre- or post-treatment with AFP-L3. Glycan microarrays showed that the AFP-L3 ratio of HBV-related HCC patients was significantly higher than that measured for chronic hepatitis B patients. Overall parameters for estimating AFP-L3% in HCC samples were as follows: sensitivity, 53.13%; specificity, 88.89%; and area under the curve, 0.75. The elevated AFP-L3% in the 8 patients with HBV-related HCC was strongly associated with HCC progression. Following one month of transarterial embolization therapy, the relative mean AFP-L3% decreased significantly. In addition, we compared Fut8 gene expression between paired tumor and non-tumor tissues from 24 patients with HBV-related HCC. The Fut8 mRNA expression was significantly increased in tumorous tissues in these patients than that in non-tumor tissue controls. Higher expression of Fut8 mRNA in tumorous tissues in these patients was associated with poor differentiation than well and moderate differentiation. Our results describe a new glycan microarray for the sensitive and rapid quantification of fucosylated AFP; this method is potentially applicable to screening changes in AFP-L3 level for assessment of HCC progression.
authors.affiliation: The Ph.D. Program for Cancer Biology and Drug Discovery, China Medical University, Taichung, Taiwan; Genomics Research Center, Academia Sinica, Taipei, Taiwan.
authors.forename: Chen-Shiou
authors.initials: CS
authors.lastname: Wu
authors.affiliation: Graduate Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan; Department of Medicine, Chung Shan Medical University, Taichung, Taiwan; Division of Gastroenterology, Taichung Veterans General Hospital, Taichung, Taiwan.
authors.forename: Teng-Yu
authors.initials: TY
authors.lastname: Lee
authors.affiliation: The Ph.D. Program for Cancer Biology and Drug Discovery, China Medical University, Taichung, Taiwan; Graduate Institute of Cancer Biology, and Center for Molecular Medicine, China Medical University, Taichung, Taiwan; Department of Biotechnology, Asia University, Taichung, Taiwan.
authors.forename: Ruey-Hwang
authors.initials: RH
authors.lastname: Chou
authors.affiliation: Graduate Institute of Clinical Medicine, National Cheng Kung University, Tainan, Taiwan; Division of Hematology/Oncology, Department of Internal Medicine, National Cheng Kung University Hospital, Tainan, Taiwan.
authors.forename: Chia-Jui
authors.initials: CJ
authors.lastname: Yen
authors.affiliation: The Ph.D. Program for Cancer Biology and Drug Discovery, China Medical University, Taichung, Taiwan; Graduate Institute of Cancer Biology, and Center for Molecular Medicine, China Medical University, Taichung, Taiwan; Department of Biotechnology, Asia University, Taichung, Taiwan.
authors.forename: Wei-Chien
authors.initials: WC
authors.lastname: Huang
authors.affiliation: The Ph.D. Program for Cancer Biology and Drug Discovery, China Medical University, Taichung, Taiwan; Genomics Research Center, Academia Sinica, Taipei, Taiwan.
authors.forename: Chung-Yi
authors.initials: CY
authors.lastname: Wu
authors.affiliation: The Ph.D. Program for Cancer Biology and Drug Discovery, China Medical University, Taichung, Taiwan; Graduate Institute of Cancer Biology, and Center for Molecular Medicine, China Medical University, Taichung, Taiwan; Department of Biotechnology, Asia University, Taichung, Taiwan.
authors.forename: Yung-Luen
authors.initials: YL
authors.lastname: Yu
mesh_terms: D000328:Adult; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000175:diagnosis* / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay; D006515:Hepatitis B virus / Q000472:pathogenicity*; D006801:Humans; D008113:Liver Neoplasms / Q000097:blood / Q000175:diagnosis* / Q000821:virology*; D046228:Microarray Analysis / Q000379:methods*; D011134:Polysaccharides / Q000032:analysis*; D000509:alpha-Fetoproteins / Q000378:metabolism*
chemical_list: D011134:Polysaccharides; D000509:alpha-Fetoproteins