Zeng Ye, Du Xiaoqiang, Jiang Wenli, Qiu Yan
Institute of Biomedical Engineering, West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu, Sichuan, China.
Methods Mol Biol. 2022;2475:79-96. doi: 10.1007/978-1-0716-2217-9_5.
Vascular endothelial growth factor (VEGF) plays a vital role in angiogenesis, and is also involved in tumor cell growth and immunosuppression, showing very complex roles. VEGF-exosomes are released by tumor endothelial cells (ECs) following anti-angiogenesis therapies (AATs). Transwell assays enable the detection of migration and invasion capacities of tumor cells. Matrigel assays are used to evaluate the angiogenesis capacities of ECs. Here we describe the detection of VEGF content in exosomes by nano-flow cytometry, enzyme-linked immunosorbent assay (ELISA), and western blotting, and demonstrate the procedure for detection of the colon formation of tumor cells induced by exosomes, the angiogenesis of tumor cells co-cultured with ECs, the angiogenesis of tumor cells induced by exosomes in Matrigel assay in vitro and tumor xenografts.
血管内皮生长因子（VEGF）在血管生成中起关键作用，还参与肿瘤细胞生长和免疫抑制，作用非常复杂。抗血管生成疗法（AATs）后，肿瘤内皮细胞（ECs）会释放VEGF外泌体。Transwell实验可检测肿瘤细胞的迁移和侵袭能力。基质胶实验用于评估内皮细胞的血管生成能力。在此，我们描述了通过纳米流式细胞术、酶联免疫吸附测定（ELISA）和蛋白质印迹法检测外泌体中VEGF含量的方法，并展示了检测外泌体诱导肿瘤细胞结肠形成、与内皮细胞共培养的肿瘤细胞血管生成、体外基质胶实验中外泌体诱导肿瘤细胞血管生成以及肿瘤异种移植的程序。
