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. 1999 Dec;81(7):1188–1195. doi: 10.1038/sj.bjc.6690828

Glycan composition of serum alpha-fetoprotein in patients with hepatocellular carcinoma and non-seminomatous germ cell tumour

P J Johnson 1, T C W Poon 1, N M Hjelm 2, C S Ho 2, S K W Ho 1, C Welby 1, D Stevenson 3, T Patel 3, R Parekh 3
PMCID: PMC2374329  PMID: 10584881

Abstract

Although estimation of serum alpha-fetoprotein (AFP) is widely used in the diagnosis of hepatocellular carcinoma (HCC) and non-seminomatous germ cell tumours (NSGCT), the clinical usefulness of this test is limited by a low specificity. However, there exist glycoforms of AFP which may be more specific for particular tumours. Previously, detailed analysis has been prevented by the low levels of AFP in human serum. We report here the application of fluorescence labelling, sequential exoglycosidase digestion, high-performance liquid chromatography and matrix-assisted laser desorption ionization in time-of-flight mass spectrometry, to determine the glycan structures of purified serum AFP from patients with HCC and NSGCT. Eleven major glycans were found, of which seven were N-linked, and four were O-linked, to the protein backbone. The structure of the N-linked glycans (all of bi-antennary complex-type with varying degrees of sialylation, fucosylation and galactosylation) were consistent with those previously reported. The O-linked glycans (three mucin O-GalNAc type glycans with variable degrees of sialylation, one O-HexNAc monosaccharide glycan) have not previously been reported. The finding of mucin O-GalNAc type glycans was supported by the prediction of potential O-GalNAc glycosylation sites on the protein backbone by analysis of the AFP structure by molecular modelling. With knowledge of these structures it may be possible to develop more specific assays for the detection of HCC and NSGCT. © 1999 Cancer Research Campaign © 1999 Cancer Research Campaign

Keywords: hepatocellular carcinoma, non-seminomatous germ cell tumour, alpha-fetoprotein, purification, protein structure, glycosylation

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Selected References

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  1. Alpert E., Drysdale J. W., Isselbacher K. J., Schur P. H. Human -fetoprotein. Isolation, characterization, and demonstration of microheterogeneity. J Biol Chem. 1972 Jun 25;247(12):3792–3798. [PubMed] [Google Scholar]
  2. Aoyagi Y., Suzuki Y., Igarashi K., Saitoh A., Oguro M., Yokota T., Mori S., Nomoto M., Isemura M., Asakura H. The usefulness of simultaneous determinations of glucosaminylation and fucosylation indices of alpha-fetoprotein in the differential diagnosis of neoplastic diseases of the liver. Cancer. 1991 May 1;67(9):2390–2394. doi: 10.1002/1097-0142(19910501)67:9<2390::aid-cncr2820670928>3.0.co;2-v. [DOI] [PubMed] [Google Scholar]
  3. Bigge J. C., Patel T. P., Bruce J. A., Goulding P. N., Charles S. M., Parekh R. B. Nonselective and efficient fluorescent labeling of glycans using 2-amino benzamide and anthranilic acid. Anal Biochem. 1995 Sep 20;230(2):229–238. doi: 10.1006/abio.1995.1468. [DOI] [PubMed] [Google Scholar]
  4. Buamah P. K., Harris R., James O. F., Skillen A. W. Lentil-lectin-reactive alpha-fetoprotein in the differential diagnosis of benign and malignant liver disease. Clin Chem. 1986 Nov;32(11):2083–2084. [PubMed] [Google Scholar]
  5. Burditt L. J., Johnson M. M., Johnson P. J., Williams R. Detection of hepatocellular carcinoma-specific alpha-fetoprotein by isoelectric focusing. Cancer. 1994 Jul 1;74(1):25–29. doi: 10.1002/1097-0142(19940701)74:1<25::aid-cncr2820740106>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  6. Du M. Q., Hutchinson W. L., Johnson P. J., Williams R. Differential alpha-fetoprotein lectin binding in hepatocellular carcinoma. Diagnostic utility at low serum levels. Cancer. 1991 Jan 15;67(2):476–480. doi: 10.1002/1097-0142(19910115)67:2<476::aid-cncr2820670226>3.0.co;2-8. [DOI] [PubMed] [Google Scholar]
  7. Hansen J. E., Lund O., Engelbrecht J., Bohr H., Nielsen J. O., Hansen J. E. Prediction of O-glycosylation of mammalian proteins: specificity patterns of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase. Biochem J. 1995 Jun 15;308(Pt 3):801–813. doi: 10.1042/bj3080801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hansen J. E., Lund O., Tolstrup N., Gooley A. A., Williams K. L., Brunak S. NetOglyc: prediction of mucin type O-glycosylation sites based on sequence context and surface accessibility. Glycoconj J. 1998 Feb;15(2):115–130. doi: 10.1023/a:1006960004440. [DOI] [PubMed] [Google Scholar]
  9. Ho S., Cheng P., Yuen J., Chan A., Leung N., Yeo W., Leung T., Lau W. Y., Li A. K., Johnson P. J. Isoelectric focusing of alphafetoprotein in patients with hepatocellular carcinoma--frequency of specific banding patterns at non-diagnostic serum levels. Br J Cancer. 1996 Apr;73(8):985–988. doi: 10.1038/bjc.1996.192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Javadpour N. The role of biologic tumor markers in testicular cancer. Cancer. 1980 Apr 15;45(7 Suppl):1755–1761. [PubMed] [Google Scholar]
  11. Johnson P. J., Ho S., Cheng P., Chan A., Leung T., Yuen J. Germ cell tumors express a specific alpha-fetoprotein variant detectable by isoelectric focusing. Cancer. 1995 Apr 1;75(7):1663–1668. doi: 10.1002/1097-0142(19950401)75:7<1663::aid-cncr2820750717>3.0.co;2-o. [DOI] [PubMed] [Google Scholar]
  12. Johnson P. J., Leung N., Cheng P., Welby C., Leung W. T., Lau W. Y., Yu S., Ho S. 'Hepatoma-specific' alphafetoprotein may permit preclinical diagnosis of malignant change in patients with chronic liver disease. Br J Cancer. 1997;75(2):236–240. doi: 10.1038/bjc.1997.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnson P. J., Portmann B., Williams R. Alpha-fetoprotein concentrations measured by radioimmunoassay in diagnosing and excluding hepatocellular carcinoma. Br Med J. 1978 Sep 2;2(6138):661–663. doi: 10.1136/bmj.2.6138.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnson P. J., Williams R. Cirrhosis and the aetiology of hepatocellular carcinoma. J Hepatol. 1987 Feb;4(1):140–147. doi: 10.1016/s0168-8278(87)80021-1. [DOI] [PubMed] [Google Scholar]
  15. Kay P. H., Wells F. C., Goldstraw P. A multidisciplinary approach to primary nonseminomatous germ cell tumors of the mediastinum. Ann Thorac Surg. 1987 Dec;44(6):578–582. doi: 10.1016/s0003-4975(10)62138-8. [DOI] [PubMed] [Google Scholar]
  16. Kew M. C., Popper H. Relationship between hepatocellular carcinoma and cirrhosis. Semin Liver Dis. 1984 May;4(2):136–146. doi: 10.1055/s-2008-1040653. [DOI] [PubMed] [Google Scholar]
  17. Lange P. H., Fraley E. E. Serum alpha-fetoprotein and human chorionic gonadotropin in the treatment of patients with testicular tumors. Urol Clin North Am. 1977 Oct;4(3):393–406. [PubMed] [Google Scholar]
  18. Lok A. S., Lai C. L. alpha-Fetoprotein monitoring in Chinese patients with chronic hepatitis B virus infection: role in the early detection of hepatocellular carcinoma. Hepatology. 1989 Jan;9(1):110–115. doi: 10.1002/hep.1840090119. [DOI] [PubMed] [Google Scholar]
  19. Nichols C. R., Saxman S., Williams S. D., Loehrer P. J., Miller M. E., Wright C., Einhorn L. H. Primary mediastinal nonseminomatous germ cell tumors. A modern single institution experience. Cancer. 1990 Apr 1;65(7):1641–1646. doi: 10.1002/1097-0142(19900401)65:7<1641::aid-cncr2820650731>3.0.co;2-u. [DOI] [PubMed] [Google Scholar]
  20. Nomura F., Ohnishi K., Tanabe Y. Clinical features and prognosis of hepatocellular carcinoma with reference to serum alpha-fetoprotein levels. Analysis of 606 patients. Cancer. 1989 Oct 15;64(8):1700–1707. doi: 10.1002/1097-0142(19891015)64:8<1700::aid-cncr2820640824>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  21. Okuda K. Early recognition of hepatocellular carcinoma. Hepatology. 1986 Jul-Aug;6(4):729–738. doi: 10.1002/hep.1840060432. [DOI] [PubMed] [Google Scholar]
  22. Patel T., Bruce J., Merry A., Bigge C., Wormald M., Jaques A., Parekh R. Use of hydrazine to release in intact and unreduced form both N- and O-linked oligosaccharides from glycoproteins. Biochemistry. 1993 Jan 19;32(2):679–693. doi: 10.1021/bi00053a037. [DOI] [PubMed] [Google Scholar]
  23. Sheu J. C., Sung J. L., Chen D. S., Yang P. M., Lai M. Y., Lee C. S., Hsu H. C., Chuang C. N., Yang P. C., Wang T. H. Growth rate of asymptomatic hepatocellular carcinoma and its clinical implications. Gastroenterology. 1985 Aug;89(2):259–266. doi: 10.1016/0016-5085(85)90324-5. [DOI] [PubMed] [Google Scholar]
  24. Shimizu K., Katoh H., Yamashita F., Tanaka M., Tanikawa K., Taketa K., Satomura S., Matsuura S. Comparison of carbohydrate structures of serum alpha-fetoprotein by sequential glycosidase digestion and lectin affinity electrophoresis. Clin Chim Acta. 1996 Oct 15;254(1):23–40. doi: 10.1016/0009-8981(96)06369-3. [DOI] [PubMed] [Google Scholar]
  25. Smith C. J., Kelleher P. C. Alpha-fetoprotein molecular heterogeneity. Physiologic correlations with normal growth, carcinogenesis and tumor growth. Biochim Biophys Acta. 1980 Mar 12;605(1):1–32. doi: 10.1016/0304-419x(80)90020-7. [DOI] [PubMed] [Google Scholar]
  26. Taketa K., Endo Y., Sekiya C., Tanikawa K., Koji T., Taga H., Satomura S., Matsuura S., Kawai T., Hirai H. A collaborative study for the evaluation of lectin-reactive alpha-fetoproteins in early detection of hepatocellular carcinoma. Cancer Res. 1993 Nov 15;53(22):5419–5423. [PubMed] [Google Scholar]
  27. Yamashita K., Hitoi A., Tsuchida Y., Nishi S., Kobata A. Sugar chain of alpha-fetoprotein produced in human yolk sac tumor. Cancer Res. 1983 Oct;43(10):4691–4695. [PubMed] [Google Scholar]
  28. Yamashita K., Taketa K., Nishi S., Fukushima K., Ohkura T. Sugar chains of human cord serum alpha-fetoprotein: characteristics of N-linked sugar chains of glycoproteins produced in human liver and hepatocellular carcinomas. Cancer Res. 1993 Jul 1;53(13):2970–2975. [PubMed] [Google Scholar]
  29. Yoshima H., Mizuochi T., Ishii M., Kobata A. Structure of the asparagine-linked sugar chains of alpha-fetoprotein purified from human ascites fluid. Cancer Res. 1980 Nov;40(11):4276–4281. [PubMed] [Google Scholar]

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