Effects of double targeting gene therapy by suicide gene combined with endostatin gene in treatment of bladder cancer:an experimental study in vitro

Abstract

Objective To investigate the effect of double targeting gene therapy by using recom-binant adeno-associated virus-thymidine kinase (TK)-intemal ribosome entry site (IRES)-endostatin (ES) (rAAV-TIE) in vitro. Methods (1) Bladder cancer cells of T24 were cultured and transfeeted by rAAV-EGFP,and the virus transfeet effieaey was determined. (2) T24 and HUVEC were euhured and transfeeted by rAAV-ES, rAAV-MCS (blank virus), and rAAV-TIE respeetively. Seventy-two h later the levels of ES in the supernatants were measured by ELISA, and annexin V apoptosis test kit was used to examine the apoptosis. Results (1) Seventy-two h after transfeetion, ES could be detected in the super-natants of the T24 cells transfected with rAAV-ES,and rAAV-TIE. (2) The apoptosis rate of the T24 cells transfected with rAAV-TK and rAAV-TIE was 34.12% and 36.91% respectively,significantly higher than that of the T24 cells transfected with rAAV-MCS and of the eontml group (3.08% and 0.84% ,all P ＜0. 05). Conclusion rAAV-TIE inhibits the tumorigenesis and angiogenesis in bladder cancer effectively. Double targeting gene therapy against bladder cancer can be achieved by using rAAV. Key words: Bladder neoplasms; Suicide gene; Endostatin; Adeno associated vires; Gene tberapy