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.
Keywords: Glycan biomarker; Hepatocellular carcinoma; Transferrin.