Mechanism of tumor growth suppression, tumor regression and induction of dormancy by endostatin, new antiangiogenic inhibitor.

新型抗血管生成抑制剂内皮抑素抑制肿瘤生长、肿瘤消退和诱导休眠的机制。

基本信息

批准号：
12671146
负责人：
YAMAGUCHI Noriko
金额：
$ 2.43万
依托单位：
Tokyo Medicai and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671146/
关键词：
Angiogenesis Endostatin Tumor Cancer Collagen

项目摘要

Endostatin is a fragment derived from type XVIII collagen which constitutes blood vessel basement membrane, showed strong antiangiogenic activity and suppressed tumor growth. The phase II clinical study of Endostatin is performing as anti-cancer drug in USA. However, functional mechanism of endostain/collagen XVIII is still unknown. Our research goal is to analyze molecular mechanisms of action of endostatin on suppression of tumor growth, invasion and metastasis. In parallel, we estimated the efficacy of endostatin in tumor animal model.1. For isolation of endostatin receptor, the binding fraction was prepared by edostatin-immobilized affinity column. A couple of proteins in the fraction were subjected to amino acid sequencing, but we cold not obtain any data. Now we are preparing peptide fragments by enzyme digestion and analyzing amino acid sequence of these fragments.2. Endostain showed inhibitory activity of migration of endothelial cell induced by growth factor, VEGF. It is not clear the affect of endostain in signal transduction passway induced by VEGF. But we found that endostatin itself induced weak phosphorylation of MAP kinase and planed to investigate the competition of MAP kinase activation.3. To localize the active site of endostatin, mutants having amino acid substitution were prepared and compared to wild type. The active site was successfully localized the region of molecular surface. The synthetic peptide covered the region failed to show comparable activity of endostain. The new peptides covering slightly different region are prepared and the possibility of structural contribution to activity is examined.4. In animal model that has brain tumor, the recombinant endostatin regress the tumor volume to one fourth. The approach of gene therapy using plasmid DNA coding endostain was launched.

内接抑制素是源自第十一型胶原蛋白的片段，构成血管基底膜，表现出强大的抗血管生成活性并抑制肿瘤的生长。内接抑素的II期临床研究在美国作为抗癌药物进行。然而，内接/胶原蛋白XVIII的功能机制仍然未知。我们的研究目的是分析内接抑制素对抑制肿瘤生长，侵袭和转移的作用的分子机制。同时，我们估计了内接抑素在肿瘤动物模型中的功效1。为了隔离内接抑制素受体，结合级分是通过伊直生蛋白 - 毫米纤维化的亲和力柱制备的。馏分中的几个蛋白质进行了氨基酸测序，但我们冷却没有获得任何数据。现在，我们正在通过酶消化和分析这些片段的氨基酸序列来制备肽片段。2。内接位显示由生长因子诱导的内皮细胞迁移的抑制活性，VEGF。目前尚不清楚vegf诱导的信号传递通道中的内接元素的影响。但是我们发现，内接抑制素本身诱导了MAP激酶的弱磷酸化，并计划研究MAP激酶激活的竞争。3。为了定位内静脉素的活跃部位，制备了具有氨基酸取代的突变体并将其与野生型进行比较。活性位点成功地定位了分子表面的区域。覆盖该区域的合成肽未能显示出内柱的可比活性。准备了覆盖略有不同区域的新肽，并检查了对活动的结构贡献的可能性4。在具有脑肿瘤的动物模型中，重组内接抑制素将肿瘤体积恢复到四分之一。启动了使用质粒DNA编码底体的基因治疗方法。