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血管内皮生长因子(vascular endothelial growth factor,VEGF)可诱导血管生成,对肿瘤的生长和转移至关重要。目前,VEGF已成为新药研发的主要靶标,这类新药可通过阻断微血管的形成和生长达到抑制肿瘤细胞的生长或是维持肿瘤处于休眠期的目的。该研究制备了中和性VEGF单克隆抗体(monoclonal antibody,Mc Ab)并探讨其对肺癌裸鼠模型血管生成的抑制作用。利用可编码VEGF全长蛋白的重组质粒免疫Balb/c小鼠,获得中和性VEGF Mc Ab,并利用VEGF±VEGF Mc Ab诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,Hu VEC)。通过Western blot法检测VEGFR-2磷酸化水平(活化状态),进而测定VEGF Mc Ab的中和性。VEGF Mc Ab可以抑制VEGF对VEGFR-2酪氨酸磷酸化的激活作用。该研究将A549肺癌细胞接种于裸鼠构建肿瘤模型,探讨了VEGF Mc Ab抗血管生成的效果。结果表明,50μg VEGF Mc Ab治疗组动物模型的瘤重抑制率为51.5%,瘤体抑制率为62.5%。VEGF Mc Ab不仅可以抑制VEGF对人脐静脉内皮细胞VEGFR-2介导的信号转导作用,且可有效抑制小鼠肺癌模型中肿瘤的生长。
Vascular endothelial growth factor (VEGF) can induce angiogenesis, which is crucial for tumor growth and metastasis. At present, VEGF has become the main target for the development of new drugs. These new drugs can inhibit the growth of tumor cells or maintain the dormant stage of the tumor by blocking the formation and growth of microvessels. In this study, a neutralizing VEGF monoclonal antibody (McAb) was prepared and its inhibitory effect on the angiogenesis of lung cancer in nude mice was explored. Balb / c mice were immunized with a recombinant plasmid encoding the full-length VEGF protein to obtain neutralizing VEGF Mc Ab, and human umbilical vein endothelial cells (Hu VEC) were induced using VEGF ± VEGF Mc Ab. The level of VEGFR-2 phosphorylation (activation state) was detected by Western blot and the neutralization of VEGF Mc Ab was determined. VEGF Mc Ab can inhibit the activation of VEGFR-2 tyrosine phosphorylation by VEGF. In this study, A549 lung cancer cells were inoculated into nude mice to establish a tumor model to investigate the anti-angiogenic effect of VEGF Mc Ab. The results showed that the tumor weight inhibition rate of the animal model treated with 50 μg VEGF Mc Ab was 51.5% and the tumor inhibition rate was 62.5%. VEGF Mc Ab can not only inhibit the VEGF VEGF-2-mediated signal transduction of human umbilical vein endothelial cells, and can effectively inhibit tumor growth in mouse lung cancer model.