N- and O-glycome analysis of serum and urine from bladder cancer patients using a high-throughput glycoblotting method

Purpose: To develop novel diagnostic biomarkers for Bladder Cancer (BC), we concurrently evaluated serum and urine glycans in BC patients by utilizing a recently established glycoblotting method. Experiments: N- and O-glycan levels in whole serum from 45 and 13 male patients diagnosed with BC were analyzed. As a control, 29 and 10 patients with benign prostatic hyperplasia (BPH) were also studied and the results were compared. Furthermore, urine N- and O-glycome from 8 patients with muscle-invasive BC and 11 with BPH were analyzed. In serum N-glycome analysis the area-under-the-curve (AUC) value was calculated by in house R software. In the other cases JMP, version 10.0.2 software package (SAS, Cary, NC) was used and p <0.05 was considered statistically significant. Results: The expression level of three N-glycans significantly increased in sera of BC patients. All of them had highly branched and sialylated structures with core-Fuc. The levels of three O-glycans were significantly higher in BC than in the control. In examination of urine samples, 16 N-glycans were significantly elevated in BC as compared to the control. Although 11 O-glycans were detected in urine samples, there was no significant difference in the expression levels. Conclusions: The levels of highly branched, sialylated N-glycans and early sialylated O-glycans were increased in sera of BC patients. Moreover, we found that N-glycans increased in urine more than in serum of BC patients. These results suggest that the glycome change in urine directly results from glycoproteins that exist in BC cells. Thus, further large-scale glycan profiling will provide novel biomarkers for diagnosing BC in the near future.