应用重组分泌型内皮抑素腺相关病毒治疗肾细胞癌

Gene therapy of renal cell carcinoma by using rAAV-ES

【目的】研究重组分泌型内皮抑素腺相关病毒(rAAV-Endostatin,rAAV-ES)的抗肿瘤血管生成及抑制肾细胞癌进展的作用。【方法】应用rAAV-ES转染肾癌细胞,ELISA法测定上清液中重组内皮抑素的浓度并检测其对血管内皮细胞趋化运动的抑制作用;建立裸鼠肿瘤模型,检测肾癌细胞被感染后的成瘤率及全身应用rAAV-ES后抑制肿瘤发展的作用及其毒副作用。【结果】RCC细胞感染rAAV-ES后上清液中重组内皮抑素浓度为52.67 ng/ml,对血管内皮细胞趋化运动的抑制率为38.13%;转染rAAV-ES后的肾癌细胞成瘤率为对照组的60%,体内实验证实全身应用rAAV-ES后血清中内皮抑素长期高效表达,肿瘤生长速度减慢,瘤体微血管密度变低,心脑组织检查未见缺血和其他病理改变。【结论】rAAV-ES无毒副作用,可有效地抑制肿瘤的血管生成,从而抑制肾细胞癌的发生和发展。

[Objective]To study the anti-tumor effect of rAAV-Endostatin （rAAV-ES） in renal cell carcinoma. [Methods]After RCC cells infected （MOI = 105）with rAAV-ES, endostatin concentration in the supernatant was assayed by ELISA, and the inhibition of HUVEC chemotactic movement was identified by Transwell. Its suppression effect on initiation and progression of renal cell carcinoma in vivo was verified, and the side effect was also studied after rAAV-ES generally used. [ Results] Endostatin concentration in the supernatant of RCC cells infected by rAAV-ES was 52.67ng/ml, the inhibition rate of HUVEC chemotactic movement was 38.13%. Xenograft formation rate of RCC cell infected by rAAV-ES was 60%. In vivo study showed that endostatin expression was high and stable, and rAAV-ES could inhibit tumor angiogenesis, so to the initiation and progression of renal cell carcinoma. After general use, side effect such as ischemia was not found. [Conclusion]rAAV-ES can inhibit tumor angiogenesis so it can also inhibit the formation and progression of tumors, rAAV-ES is a safe agent in gene therapy of renal cell carcinoma.