Lu Bing-xin, Han Rui-fa, Tang Yang, Han Yu-zhi, Li Sheng-zhi
Tianjin Urological Surgery Institute, Tianjin 300211, China.
Zhonghua Yi Xue Za Zhi. 2007 Mar 27;87(12):802-5.
To package recombinant adeno-associated virus-endostatin (rAAV-ES) and study its anti-tumor effect in vitro and in vivo.
rAAV-ES was packaged with co-transfection technique and transfected into the human bladder cancer cells of the line EJ. 24 h later ELISA was used to examine the concentration of ES in the supernatant. The inhibition of human umbilical veins endothelial cells (HUVECs) chemotactic movement were examined by Transwell system. Nude Balb/c mice were divided into 4 groups: (1) 5 mice were inoculated with the EJ cells transfected with rAAV-ES or rAAV-enhanced yellow fluorescence protein (rAAV-EYFP) for 3 days to the subcutaneous tissues of bilateral shoulders so as to observe the growth of tumor. (2) 24 mice were injected with rAAV-ES intramuscularly and then the serum ES was examined every 10 days since the 10 th day after the injection. (3) 36 mice were randomly subdivided into 3 equal subgroups to be injected with rAAV-ES, rAAV-EYFP, or RPMI medium, inoculated with EJ cells 2 weeks later, and then killed 50 days later to observe the size of tumor. (4) 4 healthy mice and 4 mice injected with rAAV-ES for 8 weeks were killed with their hearts and brains taken out to observe the side effects.
rAAV-ES was packaged successfully. The ES concentration in the supernatant of culture fluid of the EJ cell transfected with rAAV-ES was 54.09 ng/ml. The inhibition rate of the HUVECs chemotactic movement was 37.45%. The xenograft formation rate was 2/5 for the EJ cells transfected with rAAV-ES. The serum ES levels of the mice injected with rAAV-ES remained high. The tumor size in the mice injected with rAAV-ES was significantly smaller than those of the other groups (both P < 0.01). No pathological changes was found in the hearts and brains in the mice injected with rAAV-ES.
rAAV-ES inhibits tumor angiogenesis, and tumor formation and progression. Successful packaging of rAAV-ES has laid a foundation for gene therapy of bladder cancer.
包装重组腺相关病毒-内皮抑素（rAAV-ES），并研究其在体内外的抗肿瘤作用。
采用共转染技术包装rAAV-ES，并将其转染入EJ人膀胱癌细胞系。24小时后，用ELISA检测上清液中ES的浓度。采用Transwell系统检测人脐静脉内皮细胞（HUVECs）趋化运动的抑制情况。将裸Balb/c小鼠分为4组：（1）5只小鼠双侧肩部皮下接种经rAAV-ES或rAAV-增强型黄色荧光蛋白（rAAV-EYFP）转染的EJ细胞3天，观察肿瘤生长情况。（2）24只小鼠肌肉注射rAAV-ES，自注射后第10天起每10天检测血清ES水平。（3）36只小鼠随机分为3个相等的亚组，分别注射rAAV-ES、rAAV-EYFP或RPMI培养基，2周后接种EJ细胞，50天后处死，观察肿瘤大小。（4）处死4只健康小鼠和4只注射rAAV-ES 8周的小鼠，取出心脏和大脑观察副作用。
成功包装rAAV-ES。rAAV-ES转染的EJ细胞培养液上清中ES浓度为54.09 ng/ml。HUVECs趋化运动抑制率为37.45%。rAAV-ES转染的EJ细胞异种移植形成率为2/5。注射rAAV-ES的小鼠血清ES水平持续较高。注射rAAV-ES的小鼠肿瘤大小明显小于其他组（均P<0.01）。注射rAAV-ES的小鼠心脏和大脑未发现病理变化。
rAAV-ES抑制肿瘤血管生成、肿瘤形成和进展。rAAV-ES的成功包装为膀胱癌的基因治疗奠定了基础。
