Molecular mechanism of the interaction between signals generated by BMP and mechanical stress in osteoblasts

BMP产生的信号与成骨细胞机械应力相互作用的分子机制

• 批准号: 08307012
• 负责人: NODA Masaki
• 金额: $ 4.61万
• 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 (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08307012/
• 关键词: Bone Formation; Bone Resorption; Osteoblast; Chondrocyte; Mechanical Stress

Bone and cartilage are supporting organs of the body while they also respond to physical stimuli to maintain the skeletal tissues. This research aimed to classify the roles of mechanical stress in the regulation of skeletal cell function and, therefore, generated a clonel cell line derived from such weight bearing tissue, articular cartilage, from transgenic mice harboring SV-40 large-T antigen gene. This large T-antigen is active at 33ﾟ and inactive at 37ﾟ. To establish chondrocyte cell lines, we cultured the cells at 33ﾟ at a low FBS concentration (0.5%). The established TC-6 cell aggregates were stained positive for alcian blue and toluidine blue, indicating production of cartilage matrix. TC-6 cells also express type II collagen, aggrecan and link protein genes. Extermal stress affects cells via cell attachment machinery. TC-6 cells attachment to bacteriological dish was examined and we found that TC-6 cells can survive in these dish. Therefore, these cells are highly able to establish attachment efficiently even in non-attaching environment and are useful to study mechanical stress and signal transduction via attachment machinery as well as growth factor effects such as those of BMP.

骨和软骨是身体的支持器官，同时它们也对物理刺激做出反应以维持骨骼组织。本研究旨在对机械应力在骨骼细胞功能调节中的作用进行分类，因此，从携带SV-40大T抗原基因的转基因小鼠的承重组织关节软骨中产生了克隆细胞系。这种大的T抗原在33 ℃时有活性，在37 ℃时无活性。为了建立软骨细胞系，我们在低FBS浓度（0.5%）下在33 ℃培养细胞。所建立的TC-6细胞聚集体对阿辛蓝和甲苯胺蓝染色呈阳性，表明软骨基质的产生。TC-6细胞还表达II型胶原、聚集蛋白聚糖和连接蛋白基因。外部应激通过细胞附着机制影响细胞。通过对TC-6细胞在细菌培养皿上的附着情况进行观察，发现TC-6细胞在细菌培养皿上可以存活。因此，这些细胞即使在非附着环境中也能够高度有效地建立附着，并且可用于研究机械应力和通过附着机制的信号转导以及生长因子效应（如BMP的那些）。

项目成果

Toshiyuki Kawa-uchi: "Messenger RNA Expression of the Genes Encoding Receptors for Bone Morphogenetic Protein(BMP)and Transforming Growth Factor-β(TGF-β)in the Cells from the Posterior Longitudinal Ligament in Cervical Spine" Endocrine. 5・3. 307-314 (1996

Toshiyuki Kawa-uchi：“颈椎后纵韧带细胞中编码骨形态发生蛋白（BMP）和转化生长因子-β（TGF-β）受体的基因的信使RNA表达”内分泌5·3。 307-314（1996

Ying Liu: "Sclerotome-related helix type transcription factor(scleraxis)mRNA is expressed in osteoblasts and its level is enhanced by type-β transforming growth factor" Journal of Endocrinology. 151. 491-499 (1996)

Ying Liu：“骨片相关螺旋型转录因子（scleraxis）mRNA在成骨细胞中表达，β型转化生长因子可增强其水平”《内分泌学杂志》151. 491-499（1996）。

Shunichi Murakami: "Expression of Indiana Hedgehog in Osteoblasts and Its Posttranscriptional Regulation by TGFβ" Endocrinology. 139.(in press). (1997)

Shunichi Murakami：“Indiana Hedgehog 在成骨细胞中的表达及其 TGFβ 的转录后调节”内分泌学 139。（出版中）。

Toshiyuki Kawa-uchi: "Messenger RNA Expression of the Genes Encoding Receptors for Bone Morphogenetic Protein(BMP)and Transforming Growth Factor-β(TGF-β)in the Cells from the Posterior Longitudinal Ligament in Cervical Spine" Endocrine. 5. 307-314 (1996)

Toshiyuki Kawa-uchi：“颈椎后纵韧带细胞中骨形态发生蛋白 (BMP) 和转化生长因子-β (TGF-β) 受体基因的信使 RNA 表达”内分泌 5. 307- 314 (1996)

Toshiyuki Kawa-uchi: "Messenger RNA Expression of the Genes Encoding Receptors for Bone Morphogenetic Protein (BMP) and Transforming Growth Factor-beta (TGF-beta) in the Cells from the Posterior Longitudinal Ligament in Cervical Spine" Endocrine. 5. 307-3

Toshiyuki Kawa-uchi：“颈椎后纵韧带细胞中编码骨形态发生蛋白 (BMP) 和转化生长因子-β (TGF-β) 受体的基因的信使 RNA 表达”内分泌。