血管内皮生长因子抗体治疗骨肉瘤的实验研究

Experimental study of therapy effects of anti -VEGF antibody on osteosarcoma

目的 探讨血管内皮生长因子 (VEGF)抗体对骨肉瘤OS 732细胞株血管形成及瘤细胞生长状态的影响 ,为从抗血管生成途径治疗骨肉瘤提供依据。 方法 应用鸡胚绒毛尿囊膜模型 ,通过解剖显微镜下血管计数、原位铺片、组织切片HE染色及PCNA免疫组化和原位末端标记技术检测VEGF抗体对肿瘤血管形成、肿瘤生长的抑制作用及其对细胞增殖和凋亡的影响。 结果 VEGF抗体组血管密度 [给予抗体第 8天时为 (30 6± 4 6 ) ]明显低于对照组 [给予抗体第 8天时为 (5 1 0±6 9) ],瘤细胞较对照组减少 ,肿瘤细胞凋亡指数 (16 98± 4 81)显著高于PBS对照组 (3 75± 0 73) ,增殖指数两组差异不明显 ,同时VEGF抗体组凋亡的微血管内皮细胞增多 ,增殖期微血管内皮细胞少见。 结论 VEGF抗体能够显著抑制骨肉瘤OS 732血管形成 ,抑制内皮细胞增殖 ,促进内皮细胞凋亡 ,可能是VEGF抗体发挥抗血管生成作用的途径之一 ,并通过抑制血管形成而促进肿瘤细胞凋亡 ,最终达到抑制瘤细胞生长的作用。

Objective To investigate the effect of anti VEGF antibody on angiogenesis induced by osteosarcoma OS 732 cell line and tumor growth. Methods With a tumor model on the chick embryo chorioallantoic membrane (CAM), the inhibition of angiogenesis and tumor growth by anti VEGF antibody were observed under a stero microscope and a light microscope. Furthermore, the proliferation and apoptosis in tumor cells and endothelial cells ( EC ) were studied by TdT mediated duTP nick and labeling (TUNEL) and immunohistochemical staining using proliferating cell nuclear antigen (PCNA) monoclonal antibody. Results VEGF polyclonal antibody adminstration in tumor bearing chick embryo resulted in growth arrest of xenografts and a markedly reduction in the new capillaries which converged upon the tumor. The tumor cell apoptotic index was higher in the anti VEGF antibody treated group than the negative control group, but the proliferation index was not significantly different between them. At the same time, increased apoptosis and decreased proliferation in EC were also noted. Conclusion VEGF polyclonal antibody is able to inhibit the angiogenesis induced by OS 732 obviously, probably by the mechanism of inhibition of EC proliferation and promotion of their apoptosis, furtherly, which may contribute to the apoptosis of tumor cells and result in suppression of tumor growth.