The Effects Of Silencing Heparanase By SiRNA On Bladder Cancer BIU-87 Cells Biological Behavior And HPA's Related Genes.

Part oneThe effects of silencing heparanase by siRNA on bladder cancer BIU-87 Cells biological behavior and HPA's related genes.Section ISelection of HPA siRNA sequences and its influence on expression of HPA in bladder cancer BIU-87Objective:To design and synthesize siRNA sequences targeting HPA, and to detect its silencing effect on bladder cancer BIU-87 Cells.Methods:we had designed and synthesized four siRNA sequences targeting HPA,then transfected this siRNA into bladder cancer BIU-87 cells with LipofectamineTM2000. The mRNA and protein expression levels of HPA were detected by RT-PCR, real-time PCR and Western blot.The most effective siRNA sequence was selected.Results:Fours HPA siRNA sequences could down the expression of HPA in varying degrees. HPA-siH4 against 1496-1514 bp of encoding regions was the most effective siRNA. Compared with control groups, the mRNA and protein expression levels of HPA in BIU-87cells transfected with siRNA were decreased.Conclusion:we had successfully designed and synthesized HPA siRNA sequences. The siRNA can efficiently silence expression of HPA gene in bladder cancer BIU-87 cells. Sectionâ…¡The effects of silencing heparanase by siRNA on VEGF, MMP-9 genes and bladder cancer BIU-87 Cells biological behaviorObjective:To investigate the effects of HPA siRNA on bladder cancer BIU-87 cells biological behavior and HPA's related genes.Methods:HPA-siH4 selected by Previous research was transfected into bladder cancer BIU-87 cells with LipofectamineTM2000.The mRNA and protein expression levels of VEGF, MMP-9 were detected by RT-PCR, real-time PCR and Western blot, The cell proliferation,invasion and angiogenesis abilities of BIU-87 cells were detected by colony formation assay, transwell chamber assays and tube Formation assay respectively.Results:Compared with control cells,the protein expression levels of VEGF (P<0.05) and MMP-9 (P<0.01) in BIU-87cells transfected with HPA-siH4 were decreased.The cell proliferation (P<0.05), especially invasion (P<0.01) and angiogenesis (P<0.01) abilities of BIU-87 cells were suppressed after HPA-siH4 against 1496-1514 bp of encoding regions silencing HPA gene.Conclusion:The siRNA can efficiently silence expression of HPA gene in bladder cancer BIU-87 cells, reduce levels of VEGF and MMP-9 expression,and Significantly inhibit the malignant physiological behaviors of human bladder cancer BIU-87 cells in vitro.