Abstract
Alpha-fetoprotein (AFP) is widely used as a serological marker in the diagnosis of hepatocellular carcinoma (HCC) and non-seminomatous germ cell tumours (NSGCT). By application of isoelectric focusing (IEF) disease-specific AFP isoforms can be identified. Three major bands are apparent: + l (associated with ‘benign’ liver disease), + II (associated with HCC) and +III (associated with NSGCT). Recently, we have characterized the predominant glycans of human serum AFP and now report the application of these findings and electrospray ionization-mass spectrometry (ESI-MS) to the determination of the glycan composition of the isoforms present in the sera of 12 patients with HCC and of one patient with NSGCT. ESI-MS allowed simultaneous identification of various AFP glycoforms in purified serum AFP. Seven glycoforms were identified, but with different abundance in the sera of the HCC patients, whereas six glycoforms were identified in the serum from the NSGCT patient. The glycan structures of these glycoforms were deduced from their observed masses. AFP glycoforms carrying a single biantennary complex-type N-glycan appeared as the predominant glycoforms, whereas those carrying both N-glycan and O-glycan appeared as minor glycoforms. Correlation between the abundance of the AFP glycoforms and the IEF band intensity suggested that different degrees in sialylation cause the formation of isoforms. This contention was subsequently supported by the ESI-MS and kinetic in vitro desialylation studies on purified Bands + I and + II AFPs. Our findings indicate that HCC-associated isoforms (Band + II) represent a group of glycoproteins whose carbohydrate structures are all characterized by being mono-sialylated, whereas those associated with benign liver disease and NSGCT are di- and a-sialo species, respectively. Knowledge of the structure of the tumour-specific isoforms should form an important basis for clinically useful assays. © 2000 Cancer Research Campaign
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Aoyagi Y, Suzuki Y, Igarashi K, Saitoh A, Oguro M, Yokota T, Mori S, Suda T, Isemura M and Asakura H (1993) Carbohydrate structures of human α-fetoprotein of patients with hepatocellular carcinoma: presence of fucosylated and non-fucosylated triantennary glycans. Br J Cancer 67: 486–492
Buamah PK, Harris R, James OFW and Skillen AW (1986) Lentil-lectin-reactive alpha-fetoprotein in the differential diagnosis of benign and malignant liver disease. Clin Chem 32: 2083–2084
Burditt LJ, Johnson MM, Johnson PJ and Williams R (1994) Detection of hepatocellular carcinoma-specific alpha-fetoprotein by isoelectric focusing. Cancer 74: 25–29
Dell A, Khoo K-H, Panico M, McDowell RA, Etienne AT, Reason AJ and Morris HR (1993) FAB-MS and ES-MS of glycoproteins. In: Glycobiology, a practical approach, Fukuda M, Kobata A (eds) pp 187–222, IRL Press: Oxford
Du MQ, Hutchinson WL, Johnson PJ and Williams R (1991) Differential alpha-fetoprotein lectin binding in hepatocellular carcinoma. Cancer 67: 476–480
Ferranti P, Pucci P, Marino G, Fiume I, Terrana B, Ceccarini C and Malorni A (1995) Human α-fetoprotein produced from Hep G2 cell line: structure and heterogeneity of the oligosaccharide moiety. J Mass Spectr 30: 632–638
Ho S, Cheng P, Chan A, Leung N, Yeo W, Leung T, Lau WY, Li AKC and Johnson PJ (1996) Isoelectric focusing of alpha-fetoprotein in patients with hepatocellular carcinoma – frequency of specific banding patterns at non-diagnostic serum levels. Br J Cancer 73: 985–988
Javadpour N (1980) The role of biologic tumour markers in testicular cancer. Cancer 45: 1755–1761
Johnson PJ and Williams R (1987) Cirrhosis and the aetiology of hepatocellular carcinoma. J Hepatol 4: 140–147
Johnson PJ, Portmann B and Williams R (1978) Alpha-fetoprotein concentration measured by radioimmunoassay in the diagnosing and excluding of hepatocellular carcinoma. BMJ 2: 661–663
Johnson PJ, Ho S, Cheng P, Chan A, Leung T and Yuen J (1995) Germ cell tumours express a specific alpha-fetoprotein variant detectable by isoelectric focusing. Cancer 75: 1663–1668
Johnson PJ, Leung N, Cheng P, Welby C, Leung T, Lau WY and Ho S (1997) ‘Hepatoma-specific’ alpha-fetoprotein may permit preclinical diagnosis of malignant change in patients with chronic liver disease. Br J Cancer 75: 236–240
Johnson PJ, Poon TCW, Hjelm NM, Ho CS, Ho SKW, Welby C, Stevenson D, Patel T, Parekh R and Townsend RR (1999) Glycan composition of serum alpha-fetoprotein in patients with hepatocellular carcinoma and non-seminomatous germ cell tumour. Br J Cancer 81: 1188–1195
Kay PH, Wells FC and Goldstraw P (1987) A multi-disciplinary approach to primary nonseminomatous germ cell tumours of the mediastinum. Ann Thorac Surg 44: 578–582
Kew MC and Popper H (1984) Relationship between hepatocellular carcinoma and cirrhosis. Sem Liver Dis 4: 136–146
Lange PH and Fralay EE (1977) Serum AFP and HCG in the treatment of patients with testicular tumours. Urol Clin North Am 4: 393–406
Lok ASF and Lai CL (1989) Alpha-fetoprotein monitoring in Chinese patients with chronic hepatitis B virus infection: role in the early detection of hepatocellular carcinoma. Hepatology 9: 110–115
Nichols CR, Saxman S, Williams SD, Loehrer PJ, Miller ME, Wright C and Einhorn LH (1990) Primary mediastinal nonseminomatous germ cell tumours. A modern single institution experience. Cancer 65: 1641–1646
Nomura F, Ohnishi K and Tanabe Y (1985) Clinical features and prognosis of hepatocellular carcinoma with reference to serum alpha-fetoprotein levels. Cancer 64: 1700–1707
Okuda K (1986) Early recognition of hepatocellular carcinoma. Hepatology 6: 729–738
Ruoslahti E (1979) α-fetoprotein in cancer and fetal development. Adv Cancer Res 29: 275–346
Sheu JC, Sung JL, Chen DS, Yang PM, Lai MY, Lee CS, Hsu HC, Chuang CN, Yang PC and Wang TH (1985) Growth rate of asymptomatic hepatocellular carcinoma and its clinical implications. Gastroenterol 89: 259–266
Shimizu K, Katoh H, Yamashita F, Tanaka M, Tanikawa K, Taketa K, Satomura S and Matsuura S (1996) Comparison of carbohydrate structures of serum α-fetoprotein by sequential glycosidase digestion and lectin affinity electrophoresis. Clin Chim Acta 254: 23–40
Smith CJP and Kelleher PC (1980) Alpha-fetoprotein molecular heterogeneity: physiologic correlations with normal growth, carcinogenesis and tumour growth. Biochim Biophys Acta 605: 1–31
Stibler H (1991) Carbohydrate-deficient transferrin in serum: a new marker of potentially harmful alcohol consumption reviewed. Clin Chem 37: 2029–2037
Taketa K, Endo Y, Sekiya C, Tanikawa K, Koji T, Satomura S, Matsuua S, Kawai T and Hirai H (1993) A collaborative study for the elevation of lectin-reactive alpha-fetoproteins in early detection of hepatocellular carcinoma. Cancer Res 53: 5419–5423
Tarelli E, Ashcroft AE, Calam DH, Corran PH, Green BN and Wheeler SF (1992) Human alpha-fetoprotein. Molecular weight data from electrospray mass spectrometry (ESMS) analysis. Biol Mass Spectr 23: 315–317
Yamashita K, Taketa K, Nishi S, Fukushima K and Takashi O (1993) Sugar chains of human cord serum α-fetoprotein: characteristics of N-linked sugar chains of glycoproteins produced in human liver and hepatocellular carcinomas. Cancer Res 53: 2970–2975
Yoshima H, Mizuochi T, Ishii M and Kobata A (1980) Structure of the asparagine-linked sugar chains of α-fetoprotein purified from human ascites fluid. Cancer Res 40: 4276–4281
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Johnson, P., Poon, T., Hjelm, N. et al. Structures of disease-specific serum alpha-fetoprotein isoforms. Br J Cancer 83, 1330–1337 (2000). https://doi.org/10.1054/bjoc.2000.1441
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1054/bjoc.2000.1441
Keywords
This article is cited by
-
Establishment and characterization of a fucosylated α-fetoprotein-specific monoclonal antibody: a potential application for clinical research
Scientific Reports (2019)
-
Serum glycopattern and Maackia amurensis lectin-II binding glycoproteins in autism spectrum disorder
Scientific Reports (2017)
-
Carbohydrate biomarkers for future disease detection and treatment
Science China Chemistry (2010)
-
Alpha-fetoprotein (AFP)-derived peptides as epitopes for hepatoma immunotherapy: a commentary
Cancer Immunology, Immunotherapy (2009)
-
Role of sialyltransferases involved in the biosynthesis of Lewis antigens in human pancreatic tumour cells
Glycoconjugate Journal (2005)