Kibbey M C, Grant D S, Kleinman H K
Laboratory of Developmental Biology, National Institute of Dental Research, Bethesda, Md. 20892.
J Natl Cancer Inst. 1992 Nov 4;84(21):1633-8. doi: 10.1093/jnci/84.21.1633.
Angiogenesis (vascularization) has a critical role in tumor growth and metastasis, and peptides containing the SIKVAV amino acid sequence (Ser-Ile-Lys-Val-Ala-Val) have been shown to stimulate many angiogenic activities in vitro. The use of model systems to identify agents that stimulate or inhibit angiogenesis may lead to the development of new antitumor strategies.
Our purpose was to use an in vivo murine model system to study the angiogenic activity of a synthetic peptide derived from the laminin A protein chain and containing the SIKVAV amino acid sequence. We also examined the ability of the peptide to enhance tumor growth in vivo.
The SIKVAV-containing peptide was mixed with Matrigel, a reconstituted basement membrane extract used to assay stimulation of angiogenesis. The mixture was subcutaneously injected into C57BL/6 mice. At various times after injection, the Matrigel plug was excised, and angiogenic activity was assessed by histologic examination and immunohistochemical staining with an antibody to the von Willebrand factor (vWF), an endothelium-specific antigen. In other experiments, the mixture of peptide and Matrigel was co-injected with B16F10 murine melanoma cells into C57BL/6 mice, and the resultant tumors were assessed for size and vascularization.
When co-injected with Matrigel at doses as low as 10 micrograms, the SIKVAV-containing peptide stimulated angiogenesis fourfold greater than that seen in controls, and maximum angiogenic activity was observed 2 weeks after injection. This peptide was angiogenic in a dose-dependent manner up to a 100-micrograms dose. When co-injected with Matrigel and B16F10 melanoma cells, the peptide enhanced tumor growth by approximately 2.5-fold, and tumor vascularization was significantly increased (P = .027) over that observed after injection with melanoma cells and Matrigel alone.
These data demonstrate that the laminin-derived SIKVAV-containing peptide is angiogenic in a new in vivo model system and can enhance tumor vascularization and growth.
血管生成（血管化）在肿瘤生长和转移中起关键作用，含有SIKVAV氨基酸序列（丝氨酸 - 异亮氨酸 - 赖氨酸 - 缬氨酸 - 丙氨酸 - 缬氨酸）的肽已被证明在体外可刺激多种血管生成活性。使用模型系统来鉴定刺激或抑制血管生成的药物可能会导致新的抗肿瘤策略的发展。
我们的目的是使用体内小鼠模型系统来研究源自层粘连蛋白A蛋白链且含有SIKVAV氨基酸序列的合成肽的血管生成活性。我们还研究了该肽在体内增强肿瘤生长的能力。
将含SIKVAV的肽与基质胶混合，基质胶是一种用于检测血管生成刺激的重组基底膜提取物。将混合物皮下注射到C57BL / 6小鼠体内。注射后不同时间，切除基质胶栓，并通过组织学检查和用针对血管性血友病因子（vWF）（一种内皮特异性抗原）的抗体进行免疫组织化学染色来评估血管生成活性。在其他实验中，将肽和基质胶的混合物与B16F10小鼠黑色素瘤细胞共同注射到C57BL / 6小鼠体内，并评估所得肿瘤的大小和血管化情况。
当以低至10微克的剂量与基质胶共同注射时，含SIKVAV的肽刺激血管生成的程度比对照组大四倍，并且在注射后2周观察到最大血管生成活性。该肽在高达100微克的剂量下呈剂量依赖性血管生成。当与基质胶和B16F10黑色素瘤细胞共同注射时，该肽使肿瘤生长增强约2.5倍，并且肿瘤血管化比单独注射黑色素瘤细胞和基质胶后显著增加（P = 0.027）。
这些数据表明，源自层粘连蛋白的含SIKVAV的肽在新的体内模型系统中具有血管生成作用，并且可以增强肿瘤血管化和生长。
