目的   研究以复制缺陷型重组腺病毒（adenovirus，Ad）为载体的血管内皮生长因子受体1和2（vascular endothelial growth factor receptor-1 and -2，VEGFR1R2）对新生血管的抑制作用，以达到治疗结肠肿瘤的目的。方法   利用Ad载体质粒pCA13构建携带VEGFR1R2基因的质粒，在293A细胞中表达并扩增。用蛋白质印迹法检测细胞中Ad-VEGFR1R2的表达。观察Ad-VEGFR1R2对人脐静脉内皮细胞、兔角膜新生血管、小鼠结肠癌生长和肿瘤血管生成的抑制作用。采用重复测量的方差分析对实验结果进行比较。 结果   Ad-VEGFR1R2在293A细胞内成功表达出相对分子质量为62 000的蛋白。体外实验显示其可强烈抑制静脉内皮细胞生长；体内实验表明其对兔角膜新生血管的生长具有抑制作用。结肠癌小鼠模型注射Ad-VEGFR1R2后，第32天肿瘤平均体积为（400±12） mm³，明显小于对照组〔（1 200±90） mm³〕（F=135.54，P<0.01）；与对照组相比，注射Ad-VEGFR1R2的小鼠肿瘤组织中的血管生成也较少。 结论   以复制缺陷型重组Ad为载体的VEGFR1R2能明显抑制新生血管形成，并抑制小鼠结肠癌生长。

 

 Objective   To study inhibition of vascular endothelial growth factor receptor-1 and -2 (VEGFR1R2) carried by replication-defective recombinant adenovirus (Ad) on neovascularization for the treatment of colon cancer.  Methods  Adenovirus vector carrying VEGFR1R2 gene was constructed using pCA13 plasmid, and packed and amplified by 293A cells. Western blotting was used to detect VEGFR1R2 protein expression in the infected cells. Inhibitory effects of Ad-VEGFR1R2 on venous endothelial cells, corneal neovascularization, colon cancer in a mouse model, and tumor angiogenesis were investigated. The results were compared by analysis of variance with repeated measures.  Results  Ad-VEGFR1R2-infected cells successfully expressed a protein with relative molecular mass of 62 000. In vitro experiments showed that the recombinant adenovirus strongly inhibited the growth of venous endothelial cells. In vivo experiments showed that it inhibited the inflammatory corneal neovascularization. The mouse colon cancer model had the average tumor size of (400±12) mm³ 32 days after injection of Ad-VEGFR1R2, much smaller than that in control group 〔(1 200±90) mm³〕(F=135.54, P<0.01); Ad-VEGFR1R2 group also had less angiogenesis than that of control group. Conclusion  Replication-defective recombinant adenovirus carrying VEGFR1R2 gene can significantly inhibit neovascularization and the growth of colon cancer in mice.