Construction Of RAAV-VP3Providing Anti-tumor Effect To Human Bladder Cancer In Vivo

[Objective]To assess and explore the mechanism and outcomes of gene therapy to Balb/c nude mice of human bladder cancer model by rAAV-VP3.[Methods]Gene therapy evaluation of the Balb/c nude mice model of human bladder cancer by rAAV-VP3:Bearing human bladder cancer Balb/c nude mouse xenograft model. The animals were randomly divided into four groups, including untreated group, non-tumor group, hydrochloric acid Pirarubicin treatment group, rAAV-VP3treatment group. Each group using intravesical instillation method. The anti-tumor effect of rAAV-VP3to Balb/c nude mice xenograft model of human bladder cancer was analyed by drawn the curve of tumor growth, a measure of the average tumor weight inhibition rate, calculation of the average tumor weight and histochemistry of the tumor section. The comparisons of the outcomes were performed by observation of general condition and the survival ratio in each group. Immunohistochemical staining VP3protein expression in tumor timeliness, count the positive cell percentage to compare the differential expression pattern of C-erB-2, nm23Ki-67, bcl-2, p53, C-myc, COX-2, p16, pax5, CD34, CD31, CD15between the rAAV-VP3treatment group and the hydrochloric acid Pirarubicin treatment group.[Results]Recombinant adeno-associated virus rAAV-VP3can significantly inhibit orthotopic tumor growth, higher tumor inhibition rate and can improve the survival rate of nude mice. Witch can be observed after bladder instillation of recombinant adeno-associated virus rAAV-VP3. VP3protein expression was detected by immunohistochemistry, which promote tumor apoptosis. shown recombinant adeno-associated virus rAAV-VP3inhibit the expression of Ki-67, C-erbB-2, c-myc, COX-2, CA19-9; inhibition expression of tumor vascular factor CD34and CD31; raised expression of tumor suppressor genes nm-23and p16.[Conclusions]1. BALB/c nude mice lack of inhibition and killing of mature T cell help, and thus the cellular immune function, is an ideal choice for the establishment of animal models of tumor.2. Application of cell transplantation established animal bladder cancer orthotopic model of human bladder cancer, which is in occurrence and biological behavior similar to the experimental study of human bladder cancer has a very important role.3. rAAV-VP3can significantly inhibit the growth of bladder cancer in nude mice orthotopic xenograft, no significant adverse reactions, with high security.4. VP3protein expression can be observed in tumor cells by intratumoral injection of rAAV-VP3.5. Immunohistochemical detection of VP3may inhibit Ki-67, C-erbB-2, c-myc, COX-2, CA19-9, CD34, CD31’s expression, raised tumor suppressor gene nm23. p16play the anti-tumor effect.