A Study On The Effects And Possible Mechanisms Of SERPINB3 And ADAM9 On Bladder Cancer Cell

Background: Bladder cancer(BC)is one of the most fatal type of tumor among patients with genitourinary cancer worldwide.It has been reported that429,800 new cases and 165,100 deaths from BC occur worldwide each year.About 90% bladder cancer are classified of transitional cell carcinoma.The5-year survival rate for patients with non-muscle invasive bladder cancer approaches 90 %,compared with approximately 60% for patients with muscle-invasive bladder cancer.Hence,detecting and investigating the potential mechanism contributing to BC is vital in developing novel approaches andtherapeutics.Objective: To observe the effects of SERPINB3(serineproteinase inhibitor B3)and ADAM9(adisintegrin and metalloproteinase 9)on tumorigenesis and progression of the bladder cancer.Methods: The expression of SERPINB3 gene and ADAM9 gene in the bladder cancer tissues and adjacent normal tissues was detected by the RT-PCR.ShRNA SERPINB3 and shRNA ADAM9 were constructed and transfected into bladder cancer cell line T24 with liposomes and purified from puromycin.The experiment was divided into Blank control group,negative control group,shRNA-SERPINB3 group and shRNA-ADAM9 group.The expression of SERPINB3,ADAM9,E-cadherin,Vimentin were detected by Western blot and real time real-time polymerase chain reaction(RT-PCR).The effects of shRNA SERPINB3 and shRNA ADAM9 on invasion and migration of T24 cells were observed by Transwell analysis and cell scratches.The effects of shRNA SERPINB3 and shRNA ADAM9 on proliferation of T24 cells were observed by colony formation assay and cell proliferation assay.Results: The mRNA levels of SERPINB3 gene and ADAM9 gene in the bladder cancer tissues were significantly higher than those in the adjacent normal tissues,and the level of SERPINB3 and ADAM9 expression in high-grade bladder cancer tissues were noticeably higher than low-grade bladder cancertissues.The expression levels of SERPINB3 and ADAM9 in shRNA SERPINB3 group and shRNA ADAM9 group were much lower than the blank group and NC control group.Western blot and qRT-PCR showed that the expression of E-cadherin was increased and the expression of Vimentin was decreased in the shRNA SERPINB3 group and shRNA ADAM9 group compared with blank group and negative control group.Transwell and cell scratches assay showed that SERPINB3 and ADAM9 interference inhibited migration and invasion of the cells.Colony formation assay and cell proliferation assay showed SERPINB3 and ADAM9 interference inhibited proliferation of the cells.Conclusion: SERPINB3 gene and ADAM9 gene show high expression in bladder cancer tissues and much higher in the high-grade tissues.RNA interference can effectively silence the expression of SERPINB3 and ADAM9 in bladder cancer cell line T24 and lead to the inhibition of EMT,migration,invasion,proliferation.Down-regulation of SERPINB3 inhibited the expression of ADAM9 in T24 cell lines.