On the analysis and regulatory mechanism of genes responsible for mechanical stress-induced osteoblast differentiation and osteogenesis

机械应力诱导成骨细胞分化和成骨基因分析及调控机制

• 批准号: 12470389
• 负责人: KAWASHIMA Hiroyuki
• 金额: $ 9.34万
• 依托单位: Niigata 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-12470389/
• 关键词: Tensile stress; Osteoblast differentiation; Osteogenesis; BMP - 4; Tetranectin; α-adaptin C; α-adaptinC; sIL-1RAcP; α-adaptin C; 網羅的遺伝子クローニング

Neonatal mouse calvarial sutures were cultured under tensile stress (TS) and genes either upregulated or downregulated were analyzed by RT-PCR and RAP-PCR. BMP-4, α-adaptinC, tetranectin and an novel gene, designated as J9, were among many of affected genes. BMP-4 was induced in mesenchymal fibroblastic cells, which later differentiated into preosteoblasts and osteoblasts. Whereas the role of BMP-4 in osteoblast differentiation is well known, those of other three genes were not. In our study, teteranectin expression was transiently upregulated during osteoblast differentiation and diminished before calcification took place. J9 expression gradually increased with osteoblast differentiation. Very recently, tetranectin-null mice were reported to develop spinal deformity. We also identified J9 as a soluble form of IL-1 receptor associated protein, which supposed to inhibit the IL-1 signal transduction, thereby may affect apoptosis of osteoblasts. We also confirmed that J9 increases as oste … More oblasts differentiate and mature. In situ hybridization study revealed that α-adaptinC mRNA induced in mesenchymal fibroblasts as well as preosteoblasts and osteoblasts. The induction may occur earlier than 3hr TS loading, since the protein product was also detected at 3 hr. Thus the gene appears to be upregulated even earlier than BMP-4. In contrasts to these genes, alpha adaptin C expression did not change with osteoblast differentiation. Thus α-adaptinC seems to be induced by TS per se. Alpha-adaptin C is a component of clathrin-adaptor complex AP2 and plays a role in endocytosis of molecules such as epidermal growth factor receptor (EGFR). We demonstrated that overexpression of α-adaptinC in A431 cells increased endocytosis of EGFR and that TS increased phosphorylation of ERKs, the event known to be stimulated by endocytosis of EGFR. Furthermore, extracellular potassium depletion, a milieu known to be inhibitory to endocytosis, suppressed TS-induced osteoblast differentiation. Thus all four genes seem to be positively involved in the TS-induced osteogenesis. Less

采用RT-PCR和RAP-PCR方法分析新生小鼠颅骨缝在张应力（TS）下表达上调或下调的基因。BMP-4、α-adaptinC、tetranectin和一个新的基因J 9是许多受影响的基因之一。BMP-4在间充质成纤维细胞中被诱导，其随后分化成前成骨细胞和成骨细胞。虽然BMP-4在成骨细胞分化中的作用是众所周知的，但其他三个基因的作用还不清楚。在我们的研究中，teteranectin的表达在成骨细胞分化过程中短暂上调，在钙化发生前减少。随着成骨细胞分化，J 9表达逐渐增加。最近，四连蛋白基因敲除的小鼠被报道发展成脊柱畸形。J 9蛋白是一种可溶性的IL-1受体相关蛋白，可能通过抑制IL-1信号转导，从而影响成骨细胞的凋亡。我们还证实，J 9随着骨密度的增加而增加， ...更多信息 胚细胞分化成熟。原位杂交结果显示，α-adaptinC mRNA在间充质成纤维细胞、前成骨细胞和成骨细胞中均有表达。由于蛋白质产物也在3小时检测到，因此诱导可能早于3小时TS加载发生。与这些基因相反，α适应蛋白C的表达不随成骨细胞分化而变化。因此，α-adaptinC似乎是由TS本身诱导的。α-适配蛋白C是网格蛋白-适配体复合物AP 2的组分，并且在分子如表皮生长因子受体（EGFR）的内吞作用中起作用。我们证明，α-adaptinC在A431细胞中的过表达增加了EGFR的内吞作用，TS增加了ERK的磷酸化，这是已知的EGFR内吞作用刺激的事件。此外，细胞外钾耗尽（已知抑制内吞作用的环境）抑制TS诱导的成骨细胞分化。因此，所有四个基因似乎都积极参与TS诱导的成骨作用。少

项目成果

川島博行: "Tensile stress induced osteoblast differentiation and osteogenesis in mouse calvarial suture in culture : Possible involvement of BMP-4 and other genes"J. Gravit. Physiol.. 7(2). 121-122 (2000)

Hiroyuki Kawashima：“拉伸应力诱导培养的小鼠颅骨缝中的成骨细胞分化和成骨：BMP-4 和其他基因的可能参与”J. Gravit.. 7(2) (2000)。

Ikegame M, Ishibashi O, Yoshizawa, T, Shimomura J, Komori T, Ozawa H and Kawashima H: "Tensile stress induces bone morphogenetic protein 4 in preosteoblastic and osteoblastie cells, which later differentiate into osteoblasts leading to osteogenesis in the

Ikegame M、Ishibashi O、Yoshizawa、T、Shimomura J、Komori T、Ozawa H 和 Kawashima H：“拉伸应力诱导前成骨细胞和成骨细胞中的骨形态发生蛋白 4，这些细胞随后分化为成骨细胞，导致成骨细胞中的成骨。

Kawashima H, Ikegame M, Shimomura J, Ishibashi O, Komori T, Noda T and Ozawa H: "Tensile stress induced osteoblast differentiation and osteogenesis in mouse calvarial suture in culture: Possible involvement BMP-4 and other genes."J. Gravit. Physiol.. 7. 1

Kawashima H、Ikegame M、Shimomura J、Ishibashi O、Komori T、Noda T 和 Ozawa H：“拉伸应力诱导培养的小鼠颅骨缝合处的成骨细胞分化和成骨：可能涉及 BMP-4 和其他基因。”J.

池亀美華: "Tensile stress induces bone morphogenetic protein 4 in preosteoblastic and fibroblastic cells, which later differentiate into osteoblasts leading to osteogenesis in the mouse calvariae in organ culture"J. Bone Miner. Res.. 16(1). 24-32 (2001)

Mika Ikegame：“拉伸应力诱导前成骨细胞和成纤维细胞中的骨形态发生蛋白 4，随后分化为成骨细胞，导致器官培养中的小鼠颅骨成骨”J. Bone Miner. 24-32 ( 2001）

川島博行: "Tensile stress induced osteoblast differentiation and osteogenesis in mouse calvarial suture in culture : Possible involvement BMP-4 and other genes"J.Gravit.Physiol.. 7(2). 121-122 (2000)

Hiroyuki Kawashima：“拉伸应力诱导小鼠颅骨缝合处的成骨细胞分化和成骨：可能涉及 BMP-4 和其他基因”J.Gravit.Physiol.. 7(2) (2000)。