The diagnostic value of serum DSA-TRF in hepatocellular carcinoma


TRF is a glycoprotein mainly secreted by hepatocytes, The aim of this study was to explore the diagnostic value of aberrant glycosylated serum transferrin (TRF) especially containing multi-antennary glycans in hepatocellular carcinoma (HCC).A total of 581 subjects including HCC patients, liver cirrhosis (LC) patients, chronic hepatitis (CHB) patients and healthy controls (HC) were recruited. All the subjects were randomly assigned to training group (n = 411) and validation group (n = 170). We firstly analyzed the serum protein N-glycome profiling of HCC, LC, and HC by DNA sequencer–assisted fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) technology. We established a lectin-antibody sandwich ELISA (Lectin-ELISA) method to detect multi-antennary glycans-contained TRF (DSA-TRF) in serum, in which Datura stramonium Agglutinin (DSA) was used for specific recognition. Levels of serum DSA-TRF and TRF were analyzed respectively. The diagnostic efficacies of DSA-TRF and TRF of differentiating HCC patients from CHB, LC patients and HC within the training group were evaluated using receiver operating characteristic (ROC) curve and tested in the validation group.The result found that in training group, serum TRF and DSA-TRF levels differed significantly between HCC (1.86 ± 0.50, g/L, 0.285 ± 0.06), CHB + LC (2.39 ± 0.74, g/L, 0.189 ± 0.07) and HC (1.92 ± 0.69, g/L, 0.249 ± 0.09) (HCC vs. CHB + LC, P < 0.001; HCC vs. HC, P < 0.001; CHB + LC vs. HC, P < 0.001). The area under the ROC curve (AUC) of DSA-TRF was significantly superior to AFP (0.880, 95%CI: 0.834–0.925 vs. 0.776, 95%CI: 0.725–0.827, P = 0.003) in differentiating HCC from CHB + LC. The AUC of diagnostic model GlycoTRF1 (0.981, 95%CI: 0.969–0.993) was higher than DSA-TRF and AFP alone (P<0.001) in differentiating HCC from CHB + LC, which was verified in validation group.The results indicated that the serum DSA-TRF might serve as a potential glycan biomarker for distinguishing HCC from CHB and LC.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 199

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. 1.

    Clerc, F., Reiding, K.R., Jansen, B.C., Kammeijer, G.S., Bondt, A., Wuhrer, M.: Human plasma protein N-glycosylation. Glycoconj. J. 33(3), 309–343 (2016)

  2. 2.

    Wang, M., Zhu, J., Lubman, D.M., Gao, C.: Aberrant glycosylation and cancer biomarker discovery: a promising and thorny journey. Clin. Chem. Lab. Med. 57(4), 407–416 (2019)

  3. 3.

    Drabik, A.: Glycopeptides as a Tool for Early Detection of Cancer. Proteomics Clin. Appl. 12(6), e1800108 (2018)

  4. 4.

    Wang, F.S., Fan, J.G., Zhang, Z., Gao, B., Wang, H.Y.: The global burden of liver disease: the major impact of China. Hepatology. 60(6), 2099–2108 (2014)

  5. 5.

    Mehta, A., Herrera, H., Block, T.: Glycosylation and liver cancer. Adv. Cancer Res. 126, 257–279 (2015)

  6. 6.

    Ko, Y.S., Bae, J.H., Sinn, D.H., Gwak, G.Y., Kang, W., Paik, Y.H., Choi, M.S., Lee, J.H., Koh, K.C., Paik, S.W.: The Clinical Significance of Serum Alpha-fetoprotein in Diagnosing Hepatocellular Carcinoma in a Health Screening Population. Korean J Gastroenterol. 69(4), 232–238 (2017)

  7. 7.

    Mathew, S., Ali, A., Abdel-Hafiz, H., Fatima, K., Suhail, M., Archunan, G., Begum, N., Jahangir, S., Ilyas, M., Chaudhary, A.G., Al, Q.M., Mohamad, B.S., Qadri, I.: Biomarkers for virus-induced hepatocellular carcinoma (HCC). Infect. Genet. Evol. 26, 327–339 (2014)

  8. 8.

    Cheng, J., Wang, W., Zhang, Y., Liu, X., Li, M., Wu, Z., Liu, Z., Lv, Y., Wang, B.: Prognostic role of pre-treatment serum AFP-L3% in hepatocellular carcinoma: systematic review and meta-analysis. PLoS One. 9(1), e87011 (2014)

  9. 9.

    Leerapun, A., Suravarapu, S.V., Bida, J.P., Clark, R.J., Sanders, E.L., Mettler, T.A., Stadheim, L.M., Aderca, I., Moser, C.D., Nagorney, D.M., LaRusso, N.F., de Groen, P.C., Menon, K.V., Lazaridis, K.N., Gores, G.J., Charlton, M.R., Roberts, R.O., Therneau, T.M., Katzmann, J.A., Roberts, L.R.: The utility of Lens culinaris agglutinin-reactive alpha-fetoprotein in the diagnosis of hepatocellular carcinoma: evaluation in a United States referral population. Clin. Gastroenterol. Hepatol. 5(3), 394–402, 267 (2007)

  10. 10.

    Barle, H., Nyberg, B., Essen, P., Andersson, K., McNurlan, M.A., Wernerman, J., Garlick, P.J.: The synthesis rates of total liver protein and plasma albumin determined simultaneously in vivo in humans. Hepatology. 25(1), 154–158 (1997)

  11. 11.

    Zhu, J., Lin, Z., Wu, J., Yin, H., Dai, J., Feng, Z., Marrero, J., Lubman, D.M.: Analysis of serum haptoglobin fucosylation in hepatocellular carcinoma and liver cirrhosis of different etiologies. J. Proteome Res. 13(6), 2986–2997 (2014)

  12. 12.

    Comunale, M.A., Wang, M., Anbarasan, N., Betesh, L., Karabudak, A., Moritz, E., Devarajan, K., Marrero, J., Block, T.M., Mehta, A.: Total serum glycan analysis is superior to lectin-FLISA for the early detection of hepatocellular carcinoma. Proteomics Clin. Appl. 7(9–10), 690–700 (2013)

  13. 13.

    Comunale, M.A., Lowman, M., Long, R.E., Krakover, J., Philip, R., Seeholzer, S., Evans, A.A., Hann, H.W., Block, T.M., Mehta, A.S.: Proteomic analysis of serum associated fucosylated glycoproteins in the development of primary hepatocellular carcinoma. J. Proteome Res. 5(2), 308–315 (2006)

  14. 14.

    Miyahara, K., Nouso, K., Dohi, C., Morimoto, Y., Kinugasa, H., Wada, N., Takeuchi, Y., Kuwaki, K., Onishi, H., Ikeda, F., Miyake, Y., Nakamura, S., Shiraha, H., Takaki, A., Amano, M., Nishimura, S.I., Yamamoto, K.: Alteration of N-glycan profiles in patients with chronic hepatitis and hepatocellular carcinoma. Hepatol. Res. 45(9), 986–993 (2015)

  15. 15.

    Parkkinen, J., von Bonsdorff, L., Ebeling, F., Sahlstedt, L.: Function and therapeutic development of apotransferrin. Vox Sang. 83(Suppl 1), 321–326 (2002)

  16. 16.

    Gomme, P.T., McCann, K.B., Bertolini, J.: Transferrin: structure, function and potential therapeutic actions. Drug Discov. Today. 10(4), 267–273 (2005)

  17. 17.

    Yamashita, K., Koide, N., Endo, T., Iwaki, Y., Kobata, A.: Altered glycosylation of serum transferrin of patients with hepatocellular carcinoma. J. Biol. Chem. 264(5), 2415–2423 (1989)

  18. 18.

    Matsumoto, K., Maeda, Y., Kato, S., Yuki, H.: Alteration of asparagine-linked glycosylation in serum transferrin of patients with hepatocellular carcinoma. Clin. Chim. Acta. 224(1), 1–8 (1994)

  19. 19.

    Liu, X.E., Desmyter, L., Gao, C.F., Laroy, W., Dewaele, S., Vanhooren, V., Wang, L., Zhuang, H., Callewaert, N., Libert, C., Contreras, R., Chen, C.: N-glycomic changes in hepatocellular carcinoma patients with liver cirrhosis induced by hepatitis B virus. Hepatology. 46(5), 1426–1435 (2007)

  20. 20.

    Li, L., Gu, X., Fang, M., Ji, J., Yi, C., Gao, C.: The diagnostic value of serum fucosylated fetuin A in hepatitis B virus-related liver diseases. Clin. Chem. Lab. Med. 54(4), 693–701 (2016)

  21. 21.

    DeLong, E.R., DeLong, D.M., Clarke-Pearson, D.L.: Comparing the areas under two or more correlated receiver operating characteristic curves: a nonparametric approach. Biometrics. 44(3), 837–845 (1988)

  22. 22.

    Fang, M., Zhao, Y.P., Zhou, F.G., Lu, L.G., Qi, P., Wang, H., Zhou, K., Sun, S.H., Chen, C.Y., Gao, C.F.: N-glycan based models improve diagnostic efficacies in hepatitis B virus-related hepatocellular carcinoma. Int. J. Cancer. 127(1), 148–159 (2010)

  23. 23.

    Telford, J.E., Doherty, M.A., Tharmalingam, T., Rudd, P.M.: Discovering new clinical markers in the field of glycomics. Biochem. Soc. Trans. 39(1), 327–330 (2011)

  24. 24.

    El-Aneed, A., Banoub, J.: Proteomics in the diagnosis of hepatocellular carcinoma: focus on high risk hepatitis B and C patients. Anticancer Res. 26(5A), 3293–3300 (2006)

  25. 25.

    Cui, Y., Jia, J.: Update on epidemiology of hepatitis B and C in China. J. Gastroenterol. Hepatol. 28(Suppl 1), 7–10 (2013)

  26. 26.

    Haab, B.B.: Antibody-lectin sandwich arrays for biomarker and glycobiology studies. Expert Rev Proteomics. 7(1), 9–11 (2010)

  27. 27.

    Shimomura, M., Nakayama, K., Azuma, K., Terao, N., Nishino, K., Takamatsu, S., Nakano, M., Takahashi, S., Kobayashi, Y., Murata, K., Kamada, Y., Miyoshi, E.: Establishment of a novel lectin-antibody ELISA system to determine core-fucosylated haptoglobin. Clin. Chim. Acta. 446, 30–36 (2015)

  28. 28.

    Matsumoto, H., Shinzaki, S., Narisada, M., Kawamoto, S., Kuwamoto, K., Moriwaki, K., Kanke, F., Satomura, S., Kumada, T., Miyoshi, E.: Clinical application of a lectin-antibody ELISA to measure fucosylated haptoglobin in sera of patients with pancreatic cancer. Clin. Chem. Lab. Med. 48(4), 505–512 (2010)

  29. 29.

    Wei, X., Li, N., Li, S., Shi, J., Guo, W., Zheng, Y., Cheng, S.: Hepatitis B virus infection and active replication promote the formation of vascular invasion in hepatocellular carcinoma. BMC Cancer. 17(1), 304 (2017)

Download references


This study was funded by the National Science and Technology Major Project of China (2018ZX10302205-003) and the Science and Technology Commission of Shanghai Municipality(17JC1404500, 17411960500, 15JC1404100).

Author information

Correspondence to Chunfang Gao.

Ethics declarations

Conflict of interest

The authors declare that there are no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Guan, W., Gao, Z., Huang, C. et al. The diagnostic value of serum DSA-TRF in hepatocellular carcinoma. Glycoconj J (2020).

Download citation


  • Transferrin
  • Hepatocellular carcinoma
  • Glycan biomarker