Effect Of CXCR4 Gene Silence On Invasion And Intraluminal Implantion Ability Of Human Bladder Cancer Cells

Objective:Constructed eukaryotic expression vectors of small interfering RNA targeting human CXCR4 gene and observed the expression of CXCR4 gene after the vector transfected into cells.the method of western blot tests the expression level of CXCR4 protein, the solution of RT-PCR tests EJ-M3 cells change about the expression of CXCR4 mRNA;Specific silence chemotaxis cell factor receptor CXCR4 for The influence of bladder cancer cells invasion ability and Lumen planting ability。Methods:Considering CXCR4 as target genes Using the polymerase SiRNA is synthesized in vitro according to the gene bank By the liposome 2000,the aim genes enter into bladder cancer cells, the method of western blot tests the expression level of CXCR4 protein, the solution of RT-PCR tests EJ-M3 cells change about the expression of CXCR4 mRNA, the model about Boyden small room checks the change of SiRNA-EJ-m3 invade ability, Determining cell proliferation condition by MTT method;animal model test the influence of bladder cancer cells lumen implant ability.Results:After the SiRNA genes enter into bladder cancer cells, positive cloning cells are left via G418 selection, Contrast to the blank group the protein levels of CXCR4 expression is decreased obviously (38.45%±4.4% vs 97.21%±5.33%,p<0.01);the expression level of CXCR4 mRNA (0.149±0.03) was significantly lower than control group (0.44±0.08) And negative control group (0.43±0.05), there are significant differences,(P<0.05). The invasion ability of SiRNA-EJ-m3 becomes weaker than EJ-m3 (39.67±6.45 vs 87.33±8.38 p<0.01).there is not obvious difference in the distribution of the cell cycle.but after stimulated by SDF-1,each cell proliferation increases, but SiRNA-EJ-m3 group cell proliferation is significantly lower than control group (24h,0.61±0.05 vs 0.78±0.03 P<0.05;48h 0.65±0.03 vs 0.88±0.04 P<0.05 72h P=0.80±0.04 vs 1.40±0.07 P<0.05). Animal model test group and control group contrast. Animal model is established Which tests the difference in the rate of cancer between two groups cell. (10%vs70%;P<0.05)Conclusion:The synthesized SiRNA to CXCR4 for target genes can effectively lower the expression of EJ-m3 CXCR4 genes, Can significantly inhibit the protein expression of EJ-M3 and the CXCR4 mRNA transcription,can also reduce the invasion ability and proliferation activity bladder cancer cells in vitro under induced by SDF-1.At the same time, can also lower the lumen implantion ability of EJ-m3.