Development of analytical method for each domain of CTGF/ecogenin and its clinical application.

CTGF/ecogenin各结构域分析方法的开发及其临床应用。

• 批准号: 12557154
• 负责人: TAKIGAWA Masaharu
• 金额: $ 8.45万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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/en/grant/KAKENHI-PROJECT-12557154/
• 关键词: CTGF; chondrocytes; osteoblasts; CCN family; ELISA; Cell differentiation; angiogenesis(diseases); hypoxia

Expression vectors for IGF binding protein-like(IGFBP), von Willebrand type C repeat(VWC), thrombospondin type I repeat(TSP1) and C-terminal(CT) domains of CTGF/ecogenin were constructed and their recombinant proteins were produced.Plasmids of CTGF lacking 1 or 2 domains were constructed and introduced into HeLa cells. Consequently, HeLa cell lines producing these proteins were established.Using the recombinant proteins, epitopes of six monoclonal antibodies and two polyclonal antibodies were determined. Three types of ELISA systems were developed.After production, CTGF was processed before it was secreted out of cells.CTGF was found to be a hypoxia-induced angiogenic factor, tumor angiogenesis factor and mechanical stress-induced factor. The ELISA systems described above could be used for diseases related with these phenomina.

构建了CTGF/ecogenin的IGF结合蛋白样（IGFBP）、vonWillebrand C型重复序列（VWC）、血小板反应蛋白I型重复序列（TSP 1）和C末端（CT）结构域的表达载体，并获得了重组蛋白，构建了缺失1或2个结构域的CTGF质粒，将其导入HeLa细胞。建立了分泌这些蛋白的HeLa细胞系，并利用重组蛋白对6株单克隆抗体和2株多克隆抗体的表位进行了测定。建立了三种不同类型的ELISA体系，CTGF在产生后经过处理后才被分泌到细胞外，发现CTGF是缺氧诱导的血管生成因子、肿瘤血管生成因子和机械应力诱导因子。上述ELISA系统可用于与这些现象相关的疾病。

项目成果

Kubota, S.: "Novel intracellular effects of human connective tissue growth factor expressed in Cos-7 cells"FEBS Letters. 474. 58-62 (2000)

Kubota, S.：“Cos-7 细胞中表达的人结缔组织生长因子的新细胞内效应”FEBS Letters。

Takashi Yamashiro et al.: "Mechanical stimulation induces CTGF expression in rat osteocytes. Takigawa,M.: Basic aspect of cytokines in the field of orthopaedic surgery."J. Dent. Res.. 80. 461-465 (2001)

Takashi Yamashiro 等人：“机械刺激诱导大鼠骨细胞中的 CTGF 表达。Takikawa,M.：骨科手术领域细胞因子的基本方面。”J.

Satoshi, Kubota.: "Identification of an RNA element that confers post-transcriptional repression of connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) gene: Similarities to retroviral RNA-protein interactions."Oncogene. 19.

Satoshi, Kubota.：“鉴定对结缔组织生长因子/肥大软骨细胞特异性 24 (ctgf/hcs24) 基因进行转录后抑制的 RNA 元件：与逆转录病毒 RNA-蛋白质相互作用的相似性。”癌基因。

中西 徹, 滝川 正春: "新・分子骨代謝学と骨粗鬆症"軟骨細胞の分化制御と軟骨形成. 55-68 (2001)

Toru Nakanishi，Masaharu Takikawa：“新分子骨代谢与骨质疏松症”软骨细胞分化和软骨形成的调节55-68（2001）。

Nishida,T.: "Effects of CTGF/Hcs24, a hypertrophic chondrocyte-specific gene product, on the proliferation and differentiation of osteoblastic cells in vitro"J.Cell.Physiol.. 184. 197-206 (2000)

Nishida,T.：“肥大软骨细胞特异性基因产物 CTGF/Hcs24 对体外成骨细胞增殖和分化的影响”J.Cell.Physiol.. 184. 197-206 (2000)