Goldbrunner R H, Bendszus M, Sasaki M, Kraemer T, Plate K H, Roosen K, Tonn J C
Department of Neurosurgery, University of Würzburg, Germany.
Neurosurgery. 2000 Oct;47(4):921-9; discussion 929-30. doi: 10.1097/00006123-200010000-00024.
The goal of the present study was to develop an orthotopic in vivo model for the investigation of vascular endothelial growth factor (VEGF)-dependent glioma growth and vascularization.
C6 glioma cells were infected with viruses encoding sense or antisense VEGF. Expression of the transgene was controlled by Northern blot analysis, Western blot analysis, and immunohistochemistry. Spheroids generated from both clones as well as from wild-type and mock-transfected cells were implanted in the brains of Sprague-Dawley rats. Growth and vascularization were assessed using magnetic resonance imaging after 7 and 11 days. Histology was studied using hematoxylin and eosin staining, immunohistochemistry with anti-von Willebrand staining, anti-VEGF, anti-CD8, and assessment of vessel density.
Cell proliferation, migration, and invasion in vitro were very similar in all cell clones. Sense gliomas demonstrated by far the fastest growth in vivo, with intense contrast enhancement meeting criteria for highly malignant tumors. Histological examination revealed masses of von Willebrand- and VEGF-positive tumor vessels with a high vessel density. Antisense gliomas depicted the radiological features of low-grade gliomas, with slow growth and poor vascularization, although they were highly infiltrative. Wild-type and mock-transfected gliomas demonstrated similar growth and vascularization patterns intermediate between sense and antisense gliomas. Any influence of the allogeneic response of the hosts on different tumor sizes could be excluded.
Our model elucidates glioma growth and vascularization as strongly VEGF dependent, which is consistent with human gliomas. Thus, this model is suitable for testing antiangiogenic strategies to interfere with the VEGF/VEGF receptor system, as well as for exploring VEGF-independent mechanisms using the antisense-transfected clone.
本研究的目的是建立一种原位体内模型，用于研究血管内皮生长因子（VEGF）依赖性胶质瘤的生长和血管生成。
用编码VEGF正义或反义序列的病毒感染C6胶质瘤细胞。通过Northern印迹分析、Western印迹分析和免疫组织化学来控制转基因的表达。将来自这两个克隆以及野生型和空载体转染细胞产生的球体植入Sprague-Dawley大鼠的脑内。在7天和11天后使用磁共振成像评估生长和血管生成情况。使用苏木精和伊红染色、抗血管性血友病因子染色、抗VEGF、抗CD8免疫组织化学以及评估血管密度来进行组织学研究。
所有细胞克隆在体外的细胞增殖、迁移和侵袭情况非常相似。正义胶质瘤在体内显示出迄今为止最快的生长速度，具有强烈的对比增强，符合高度恶性肿瘤的标准。组织学检查显示大量血管性血友病因子和VEGF阳性的肿瘤血管，血管密度高。反义胶质瘤表现出低级别胶质瘤的放射学特征，生长缓慢且血管生成不良，尽管它们具有高度浸润性。野生型和空载体转染的胶质瘤显示出介于正义和反义胶质瘤之间的相似生长和血管生成模式。可以排除宿主的同种异体反应对不同肿瘤大小的任何影响。
我们的模型阐明了胶质瘤的生长和血管生成强烈依赖于VEGF，这与人类胶质瘤一致。因此，该模型适用于测试干扰VEGF/VEGF受体系统的抗血管生成策略，以及使用反义转染克隆探索不依赖VEGF的机制。
