98 / 2021-10-12 08:05:50

七鳃鳗来源LIP对人尿液UMOD糖蛋白识别及其在膀胱癌检测应用

七鳃鳗，LIP，尿调素，膀胱癌，尿液

Purpose: Clinical management of bladder cancer (BCa) is hindered by the lack of reliable fbiomarkers. We aimed to investigate the value of lamprey immunity protein (LIP), a lectin that specifically binds to bi- and tri-antennary sialylated containing N-glycolylneuraminic acid (Neu5Gc) structures decorated UMOD glycoprotein in urine from BCa patients.

Experimental Design:  The UMOD protein was extracted and purified from fresh BCa urine and distributed to analyze the structure of Neu5Gc-modified glycan chains of LC/MS/MS profiling. In addition, a LIP strip method based on TRFIA was used to evaluate the accuracy of BCa screening platforms.

Results: We established dot blotting and ELISA method for LIP recognition of Neu5Gc and identified a significantly higher expression of specific Neu5Gc-modified glycan in the urine and tissue samples of BCa patients than that of the healthy participants. Primary BCa patients had higher levels of LIP-positive binding Neu5Gc in urine, compared with healthy participants and patients of postoperative treatment. In addition, lectin chip assay and mass spectrometry were used to analyze the glycan chain structure, LIP can recognize bi- and tri-antennary sialylated containing Neu5Gc structures decorated UMOD glycoprotein. The expression of sialyltransferases such as ST3Gal1 and ST3Gal4 are upregulated in bladder tumor tissues by quantitative real time RT-PCR and immunohistochemistry. Furthermore, we established a LIP strip method based on TRFIA for an initial diagnosis and monitor of BCa prognosis. Compared with heathy (N=2821, T/C=0.12±0.09) and benign urine (N=360, T/C=0.11±0.08), the range of urine T/C ratio detected by LIP test paper was 1.97±0.32 in patients with bladder cancer (N=518) with significant difference (P<0.0001).

Conclusions: Our results indicated that LIP may be a biomarker of diagnosis and monitored for early BCa identification through recognizing Neu5Gc-modified UMOD glycoprotein in urine, and Neu5Gc-modified N-glycochains and sialyltransferases act as potentially marker in clinical trials.