The Expression Of BAI-1in Bladder Cancer And The Research On Inhibition Of Angiogenesis Of Tumor

Part I The expression of BAI-1in the bladder transitional cell carcinoma tissues and the correlation between BAI-1and p53、MVDand VEGFObjective:To investigate the different expression of different stages of BTCC. To investigate the mechanism of BAI-1the inhibits the proliferation of tumor endothelial cell.Methods:Paraffin section preparation of131cases of BTCC and28cases of normal bladder mucosal tissue were taken. Normal bladder mucosal tissue section was set as control group and BTCC tissue section was set as observe group. Immunohistochemical SP method was used to detect the expression of BAI-1, VEGF, MVD and p53, and semi-quantitative statistically analysis was use.21cases of BTCC and28cases of normal bladder mucosal tissue of fresh tissue samples were taken and western blot was used to detect the different expression of BAI-1between them.Result:The result of Immunohistochemical staining and western blot showed that the expression of BAI-1in normal bladder mucosal tissue was higher than that in BTCC tissue and the expression of BAI-1was correlated to clinical stages. The expression of BAI-1in BTCC tissue of T1stage was much higher than that of T2-4(P <0.05). The expression of BAI-1had negative relationship to the expression of VEGF (r=-0.661, P=0.000). The expression of BAI-1had negative relationship to the expression of MVD (r=-0.406, P=0.002). The expression of BAI-1had negative relationship to the expression of mutant type P53(r=-0.675,P=0.000)Conclusion:The content of BAI-1in BTCC tissue is much lower than that in normal bladder mucosal tissue. The content of BAI-1is related to the clinical stage of BTCC. The expression of BAI-1had negative relationship to the expression of VEGF, MVD and mutant type P53, which shows that BAI-1may participate in the negative regulation of proliferation of tumor micrangium. The reduction of the expression of BAI-1may relate to the mutant of P53. Part II the study of recombinant plasmid vector pReceiver-M61-BAI1transfect T24and HUVECObjective:To construct and identify recombinant plasmid vector pReceiver-M61-BAI1that includes the gene of BAI-1for studying its role in human carcinoma cells of urinary bladder (T24) and human vascular endothelial cell of umbilical vein (HUVEC).Methods:Recombinant eukaryotic expression vector pReceiver-M61-BAI1was transformed and indentified. pReceiver-M61-BAI1was used to transfect T24and HUVEC and stable transfected cells were cultured. Null vector transfection was used as the control. RT-qPCR and Western blot were used to detect the expression of gene and protein. Flow cytometry was used to study the apoptosis of the T24and HUVEC before and after gene transfection. MTT was used to study the proliferation of the T24and HUVEC before and after gene transfection.Result:pReceiver-M61-BAI1identified by restriction enzyme cleave was verified including BAI-1gene. The RT-PCR product of pReceiver-M61-BAI1-T24cell and pReceiver-M61-BAI1-HUVEC cell showed obvious positive stripe by gel electrophoresis,which was not observed in the control group. The result of Western blot showed that the positive cells rate of BAI-1in pReceiver-M61-BAI1-T24cell and pReceiver-M61-BAI1-HUVEC cell were much higher than that of the control group. The result of MTT showed that the absorbance value A of the pReceiver-M61-BAI1-HUVEC cell at the second day, the third day and forth day were much lower than that of the control group and pReceiver-M61-BAI1-T24cell group. The result of flow cytometry showed that the apoptosis rate of HUVEC transfected BAI-1was much higher than that of the control group and the pReceiver-M61-BAI1-T24cell group.Conclusion:BAI-1has obviously inhibited effect on the vascular endothelial cells. While directed inhibited effect of BAI-1on T24cells is not found in vitro.