西妥昔单抗抑制血管生成的实验研究

A experimental study on inhibiting effect of cetuximab on angiogenesis

目的探讨西妥昔单抗(C225)对体内、外血管生成的抑制作用。方法用人脐静脉内皮细胞(HUVEC)原代培养模型,分别采用四甲基偶氮唑盐(MTT)法、Transwell小室趋化实验、体外小管形成实验及流式细胞术,观察C225在体外对HUVEC增殖、迁移、小管形成和凋亡的影响;利用鸡胚绒毛尿囊膜(CAM)模型,观察C225对CAM新生血管的抑制作用。结果0.0625～2.0000μg/ml的C225作用HUVEC48h,细胞增殖抑制率为12.34%～25.26%;体外迁移及小管形成实验显示,0.0625～1.0000μg/mlC225的HUVEC迁移抑制率为15.51%～48.68%,HUVEC小管形成抑制率为22.91%～44.71%;0.25μg/ml和1.00μg/ml的C225诱导HUVEC细胞凋亡率分别为33.20%和35.05%。体内CAM试验表明,0.0625～1.0000μg/ml的C225对新生血管抑制率为20.39%～36.18%。结论C225在体内、外具有抑制血管生成作用。

Objective To study the effect of cetuximab （C225） on anti-angiogenesis in vitro and in vivo. Methods Human umbilical vein endothelial cells （HUVEC） were cultured primarily in vitro. MTT colorimetric assay was used to observe the effect of C225 on the proliferations of hUVEC. Transwell migration assay and out-body tube formation test were used to observe the impact of C225 on the migration and vaso-formed ability of HUVEC, and the rate of C225 inducing hUVEC apoptosis was calculated by flow cytometry. The Chick embryo chorioallantoic （CAM） model was used to check the antiangiogenie effect of C225 in vivo. Results C225 （0. 0625- 2. 0000 μg/ml） inhibited the proliferation of hUVEC by 12.34% to 25.26% after treatment for 48 h. The inhibition rate of migration was 15.51% to 48.68% when treated with C225 （0. 0625-1. 0000μg/ml）. The inhibition rate of tube formation was 22.91% to 44.71% when treated with C225 （0. 0625-1. 0000 μg/ml）. Moreover,when hUVEC was incubated with C225 0.25 μg/ml and 1.00 μg/ml, the rates of cell apoptosis were 33.20% and 35.05%, respectively. It was observed that C225 suppressed the neovascularization of CAM in vivo with the inhibition rate of 20. 39% to 36. 18% when treated with C225 （0. 0625-1. 0000 μg/ml）. Conclusion C225 has anti angiogenic ability in vitro and in vivo.