ANALYSIS OF SIGNAL TRANSDUCTION PATHWAY IN BONE INDUCTION BY BONE MORPHOGENETIC PROTEIN

骨形态发生蛋白骨诱导信号转导通路分析

• 批准号: 09672039
• 负责人: ASAHINA Izumi
• 金额: $ 1.92万
• 依托单位: TOKYO MEDICAL 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: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672039/
• 关键词: BMP; Smad; cbfa-1; signal transduction; 情報伝達; Cbfa-1

(1) Phospholilation of mouse Smad-l by BMPMouse Smad-1 cDNA fragment of 1.6 kb was obtained from cDNA of mouse bone marrow derived stroma cell : ST-2 using PCR method, and the cDNA in protein expression vector with Flag reporter gene was transfected into ST-2 cell.. The transformed cell was treated with OP-1, BMP-2, and TGF-beta, and then cell lysate was imunoprecipitated with the antibody against Flag protein. As a result, phospholilation of mSmad was observed in the cell treated with these growth factors. These results suggest Smad play important role in the signal transudation pathway of osteoblastic differentiation.(2) Induction of cbfa-1 gene expressionWe examined the gene expression of runt family transcriptional factor, cbfa- 1, which plays important role in bone formation. The treatment of OP-1 to ST-2 or MC3T3-El cell induced induced cbfa-l gene expression two to three times by control after 48 hours. Thus, the result suggests that BMP signal is transmitted to cbfa-l byway of Smad phosphlilation and stimulates osteoblastic differentiation.

(1)BMP对小鼠Smad-1的磷酸化Mouse Smad-1 cDNA片段1.6kb是利用PCR方法从小鼠骨髓源性基质细胞ST-2的cDNA中获得的，并且将具有Flag报告基因的蛋白质表达载体中的cDNA转染入 ST-2 细胞。将转化的细胞用 OP-1、BMP-2 和 TGF-β 处理，然后将细胞裂解物用针对 Flag 蛋白的抗体。结果，在用这些生长因子处理的细胞中观察到mSmad的磷酸化。这些结果表明Smad在成骨细胞分化的信号转导途径中发挥着重要作用。(2)cbfa-1基因表达的诱导我们检测了runt家族转录因子cbfa-1的基因表达,该转录因子在骨形成中发挥着重要作用。 OP-1处理ST-2或MC3T3-E1细胞48小时后诱导的cbfa-1基因表达是对照的2至3倍。因此，结果表明BMP信号通过Smad磷酸化传递至cbfa-1并刺激成骨细胞分化。

项目成果

小山治彦: "Differential Display法を用いたOsreogenic Protein-1(OP-1)の骨形成作用に伴う細胞内シグナル伝達に関与する遺伝子の解析" 口腔病学会雑誌. 642. 205-222 (1997)

Haruhiko Koyama：“使用差异显示方法分析参与与成骨蛋白-1 (OP-1)成骨作用相关的细胞内信号转导的基因”口腔学会杂志 642. 205-222 (1997)。

小山治彦: "Differential Display 法を用いた Osteogenic Protein-1(OP-1)の骨形成作用に伴う細胞内シグナル伝達に関与する遺伝子の解析" 口腔病学会雑誌. 64. 205-222 (1997)

Haruhiko Koyama：“使用差异显示法分析参与与成骨蛋白-1 (OP-1)成骨作用相关的细胞内信号转导的基因”口腔学会杂志 64. 205-222 (1997)。

Asahina I: "Molecular mechanism of bone formation and regenaration of bone defects using bone Morphogenetic protein" Rinsyo kagaku. 34. 1261-1268 (1998)

Asahina I：“利用骨形态发生蛋白进行骨形成和骨缺损再生的分子机制”Rinsyo kagaku。

Oida,S.et al: "Biological activities of bone extract containing TGFβ super family proteins" J.Hard Tiss.Biol. 7. 27-34 (1998)

Oida, S. 等人：“含有 TGFβ 超家族蛋白的骨提取物的生物活性”J.Hard Tiss.Biol. 7. 27-34 (1998)

Oida, S., Morotome, Y., Nakamura, T.and Terashima, T: "Molecular cloning of PCR amplified BMP-related genes (GDF-5,6 and 7) from rat tooth cells using modified pBluescript SK+ vector." J.Hard Tiss.Biol. 6. 16-20 (1997)

Oida, S.、Morotome, Y.、Nakamura, T. 和 Terashima, T：“使用改良的 pBluescript SK 载体从大鼠牙齿细胞中分子克隆 PCR 扩增的 BMP 相关基因（GDF-5,6 和 7）。”