Study Of The Expression Of CDA In Bladder Urothelial Carcinoma And Its Influence Of Biological Behavior On Bladder Urothelial Carcinoma Cells

Objective1. To screen and confirm the high expressive gene which is corrolated with bladder urothelial carcinoma;2. To identify the expression of cytidine deaminase(CDA) in bladder urothelial carcinoma and analyze its relationship with clinic pathological parameters, as well as its prognostic outcomes;3. To investigate the effect of CDA on invasive and stem ability of bladder urothelial carcinoma cells.Methods4. Screen out highly invasive subclone (HM, "highly metastatic") and low invasive subclone (NM, "non-metastatic") from parent human bladder cancer cell line-5637 and test the different expressive gene in these cell lines through arrary. Confirm the target gene.5.10 pairs of bladder urothelial carcinoma tissues and normal uroepithelium were collected for RT-qPCR and Western blot to test the expression of CDA.30 bladder urothelial carcinoma tissues from patients ranged from T1 to T4 were collected for immunohistochemistry. All data were treated with statistics analysis to find out the relationship between expression of CDA and clinic pathological parameters, as well as prognosis.6. Downrergulate the expression of CDA in 5637 HM cell lines by siRNA. Western blot, transwell, flow cytometry and sphere were done to evaluate the invasive and stem ability of CDA knockdown group and control group.Results1. The result from arrary show that genes, such as CDA, were highly expressed in highly invasive bladder cancer cell lines. CDA was confirmed as the target gene.2. The expression of CDA in bladder urothelial carcinoma tissues were obviously higher than that of normal uroepithelium both in RNA and protein. The expression level of CDA were correlated to the muscle invasion of bladder cancer. The patients with high CDA expression had lower overall survival rate than those with low CDA expression.3. The invasive ability of bladder cancer cells were downregulated after the knockdown of CD A. The expression of protein markers, such as CD44,HMGA2 and SOX-2, showed a decrease trend with the knockdown of CDA, which was proved by the results of flow cytometry and sphere.Conclusion1. The high expression of CDA and its corrolation with tumor stage might illustrate that CDA, as a new marker, play some key role in the procedure of tumorigenesis and progression.2. It has been demonstrated that the knockdown of CDA can suppress the invasive and stem ability of bladder cancer cells.