Vallee B L, Riordan J F, Lobb R R, Higachi N, Fett J W, Crossley G, Bühler R, Budzik G, Breddam K, Bethune J L
Experientia. 1985 Jan 15;41(1):1-15. doi: 10.1007/BF02005853.
Angiogenesis, the process of developing a hemovascular network, is an essential feature of the growth of solid tumors, and is induced by factors secreted by tumor cells. Assay procedures suitable for the investigation of angiogenesis, and for the screening of angiogenesis factors during purification are reviewed; and a number of reports describing the purification of angiogenesis factors, primarily from the rat Walker 256 carcinoma as starting material, are discussed. Work from the authors' laboratory is also presented. Walker 256 cells grown in large-scale culture were the source of a reproducible and homogeneous source of angiogenic material. Factors secreted by these cells were isolated by a series of chromatographic steps. Ion exchange chromatography on carboxymethyl-Sephadex produced two active fractions, one of which was fractionated into several macromolecular species by lectin affinity and hydrophobic adsorption chromatography. The other gave a high mol.wt, active fraction that was resolved into a low mol.wt, active component and a non-angiogenic but possibly carrier molecule with a mol.wt of 140,000. While none of the angiogenic factors were identified chemically, the results demonstrate the existence of both high and low mol.wt tumor-secreted angiogenic substances, confirming the hypothesis for tumor-induced angiogenesis and predicting potential means to interfere with the process of tumor growth.
血管生成是形成血管网络的过程，是实体瘤生长的一个基本特征，由肿瘤细胞分泌的因子诱导产生。本文综述了适用于研究血管生成以及在纯化过程中筛选血管生成因子的检测方法；并讨论了一些主要以大鼠Walker 256癌为起始材料描述血管生成因子纯化的报告。还介绍了作者实验室的工作。在大规模培养中生长的Walker 256细胞是血管生成物质可重复且均匀的来源。这些细胞分泌的因子通过一系列色谱步骤进行分离。在羧甲基葡聚糖凝胶上进行离子交换色谱产生了两个活性级分，其中一个通过凝集素亲和色谱和疏水吸附色谱进一步分离成几个大分子物种。另一个产生了一个高分子量的活性级分，该级分被分解成一个低分子量的活性成分和一个分子量为140,000的非血管生成但可能是载体分子。虽然没有通过化学方法鉴定出任何血管生成因子，但结果表明存在高分子量和低分子量的肿瘤分泌血管生成物质，证实了肿瘤诱导血管生成的假说，并预测了干扰肿瘤生长过程的潜在方法。
