利用五特征转基因小鼠获得的全人源抗VEGF单抗的抑制血管生成及抗肿瘤活性
Anti-angiogenesis and anti-tumor activity of the fully human anti-VEGF165 monoclonal antibody obtained from the five-feature transgenic mice
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摘要: 为研究一种全人源抗VEGF单克隆抗体在体外和体内的抑制血管生成和抗肿瘤活性，利用细胞划痕实验检测抗体抑制人脐静脉内皮细胞（HUVEC）迁移的活性；MTT法检测抗体抑制分泌VEGF的肿瘤细胞增殖能力；Annexin V/PI 双染检测抗体诱导肿瘤细胞凋亡活性；通过建立胃癌原位模型，静脉注射给予不同剂量的抗体，免疫组化方法检测抗体在体内的抑制血管生成和抗肿瘤活性。结果表明一定剂量的全人源抗VEGF抗体能够显著抑制HUVEC细胞的迁移，明显抑制4种分泌VEGF的肿瘤细胞增殖并显示了一定的诱导肿瘤细胞凋亡的能力。静脉注射一定剂量的抗体能够显著抑制胃癌原位肿瘤的生长和肿瘤血管生成。进而表明该全人源抗VEGF单克隆抗体能够在体外和体内都表现出显著的抑制血管生成和抗肿瘤活性。Abstract: This study was focus on the anti-angiogenesis and anti-tumor activity both in vitro and in vivo of novel fully human anti-VEGF165 monoclonal antibody obtained from five-feature transgenic mice.The effect of human anti-VEGF mAb on migration of vascular endothelial cells was observed by scratch assay;inhibitory effect of human anti-VEGF mAb on VEGF-secreting cells was determined by MTT assay and followed by Annexin V/PI double-staining assay to detect apoptosis induced by the mAb;orthotopic gastric tumor model was established to determine anti-angiogenesis and anti-tumor activity in vivo.Our novel human anti-VEGF mAb was found dramatically inhibit HUVEC cells migration and proliferation at dose 120 μg/mL in vitro while remarkable inhibition effect on four kinds of VEGF-secreting tumor cells such as HepG2，BGC823，MCF-7，SGC-7901 tested in our study might be due to the apoptosis.After mAb was administrated at dose 10 mg/kg and 5 mg/kg for 40 days tumor inhibition was calculated as 49.9% and 27.7% respectively.And immunohistochemistry analysis of orthotopic gastric tumors confirmed significant inhibition of tumor VEGF expression and neovascularization.The fully human anti-VEGF mAb exhibited remarkable anti-angiogenesis and anti-tumor growth effects both in vitro and in vivo and it is feasible to produce therapeutic fully human monoclonal antibodies using five-feature transgenic mice.
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Keywords:
- five feature transgenic mice /
- fully human mAb /
- anti angiogenesis /
- anti tumor
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