目的： 以改良方法在鸡胚绒膜尿囊膜（chick chorioallantoic membrane，CAM）上接种人成神经胶质瘤U87细胞制备移植瘤，验证改良模型筛选抗肿瘤血管新生药物的有效性和可行性。 方法： 将鸡胚种蛋随机分为2组，分别使用传统方法与改良方法建立CAM-U87移植瘤模型；改良方法在鸡胚孵育中增加9~16 h的开窗适应期，再将特制的硅胶圈放置在CAM组织一级血管的半侧；硅胶圈内接种U87细胞建立CAM-U87移植瘤模型。模型的硅胶圈内分别加入经典抗血管新生药物沙利度胺（50、100、200 μg/ml）和自主研发抗血管新药G3B6，以DMSO为对照；体视显微镜观察肿瘤形态，H-E染色法观察瘤组织新生微血管密度 (microvessel density，MVD），原位杂交（in situ hybridization，ISH）技术检测血管标记物VEGFR2的表达。 结果： 成功制备改良CAM-U87细胞移植瘤，改良模型不影响CAM本身的血管生成，其瘤细胞接种成功率较传统模型显著升高\[（70.00±4.226）% vs (41.25±5.154)%；t=4.314，P=0.000 7\]，移植瘤体积显著增大\[（60.20±6.012）vs （15.97±2.403）mm 3；t=6.012，P<0000 1\]。沙利度胺和G3B6两药均能显著抑制移植瘤组织中的血管形成，使瘤组织中MVD显著降低\[沙利度胺200 μg/ml时（15.85±1.15）% vs （29.80±4.16）%；t=2.49，P=0.047 4\]、VEGFR2表达明显减少。 结论： 改良方法 成功制备CAM-U87移植瘤模型，经沙利度胺和G3B6验证，该模型对抗肿瘤血管新生药物的筛选具有良好的有效性和可行性。

Objective : To modify the classical chick embryo chorioallantoic membrane (CAM) xenograft model of tumuorigenesis and evaluate the effectiveness and feasibility of the modified model in the screening of anti-angiogenesis drugs. Methods: Fertilized chicken eggs randomized into two groups. Eggs in the classical model group were grafted with U87 human glioma cells and the anti-angiogenic activity of test drugs, thalidomide (50, 100, 200 μg/ml), G3B6 and DMSO (the control) was determined by following the well-established classical methods. In the modified model group, the CAM was given an additional adaptation period of 9-16 h and then grafted with U8 cells by inoculating the cells into a silicone ring and then placing the ring on the half of the grade 1 vessel of the CAM. The test drugs were added into the silicone ring where their anti-angiogenic activity was evaluated. For both groups, the tumor morphology and the microvessel density (MVD) in the tumor tissue were examined under a stereo microscope with photos taken. Changes in the tumor histology was assessed by H-E staining and the expression of VEGFR2, the marker of the angiogenesis, was assessed by in situ hybridization. Results: The success rate of the xenograft was increased significantly without any negative effect on angiogenesis in the CAM in the modified model as compared with the classical model (\[70±4.226\]% vs \[41.25±5154\]%; t=4.314, P=0.000 7). The tumor volume was also significantly increased in the modified model group as compared with thecalissical model group (\[60.20±6.012\] vs \[15.97±2.403\] mm 3; t=6.012, P<0.000 1). Both thalidomide and G3B6 effectively inhibited the angiogenesis and reduced the MVD and VEGFR2 expression in the tumor tissue. Conclusion: The modified xenograft CAM model of tumuorigenesis developed in this study seems superior over the classical CAM model in studying tumorigenesis and screening anti-angiogenesis drugs.

广东省医学科研基金资助项目（No. A2013312）；国家自然科学基金资助项目（No. 31271455，No. 31200861，No. 31100852，No. 81200308，No.31200896）