Quantification Proteomes And Glycomes Analysis Of Bladder Cancer Cells With Different Migration Potentiality

Bladder cancer(BC) is one of the most common types of human malignant cancer. Always it is devided into two types: non-muscle-invasive bladder cancer and invasive bladder cancer. So far, two bladder cancer biomarkers are clinically used, they are tissue polypeptide antigen(TPA) and carcinoembryonic antigen(CEA). They were not sufficient to diagnose the bladder cancer because they were found to have high expression level in many other cancers. So it’s necessary to find more new bladder cancer biomarkers using new technology and method.We used SILAC proteomes quantification technology to detect the differences of 3 bladder cancer cell lines(HCV29, KK47 and YTS1). The results showed that 689 proteins expressed differently with 255 proteins up-regulated and 434 proteins down-regulated. Further analysis indicated that the most common molecular function these proteins taking part in was cell binding and catalytic activity and the major biological process categorie was cellular metabolic. It was reported that the integrin, a family of transmembrane proteins, functioned in cell-to-cell and cell-to-extracellular matrix(ECM) adhesive interactions, and influence cell signaling of cell growth and differentiation. Expression of integrin α6 in three bladder cancer cell lines was quantitatively analyzed by SILAC. The ratio was 1:18:14, also the result was verified by western blotting, q PCR, histochemistry and clinical gene microarray dates. The result showed that cell line KK47 expressed the highest level of integrin α6. Furthermore, the result was confirmed in different ways. Clinical data showed the same result.EMT standing for epithelial-mesenchymal transition is reported to be a fundamental step in proliferation and metastasis of tumor cells, so it is used widely in cancer research to testify the migration ability of cells. Aberrant glycosylation of proteins was thought to have important roles in many diseases especially in progressing and invasion of cancer cells. It was reported that glycosylation changed obviously in cancer cells, and proteins with aberrant glycosylation influenced the cellular migration through changing the signal pathways. In bladder cancer, the glycosylation of molecular especially the proteins effected the invasion and migration of cancer cells.HCV29 was treated with TGFβ to build the EMT model, and the treated cell put out mesenchymal features. The expression profile of N-glycan was analyzed using lectin microarray, MALDI-TOF/TOF mass spectrometry and Glyco V4 oligonucleotide microarray. Compared to HCV29, the treated cells had higher proportion of high-mannose-type and hybrid-type N-glycan, the fucosylation level was also higher in EMT. Consistently, 5 N-glycan related genes were down-regulated.The best way to cure cancer is to detect early and cure early, so the glycoproteins expressed differently in early tumor state have the important clinical significance.