Cell-matrix interactions in bone formation and regeneration

骨形成和再生中的细胞-基质相互作用

• 批准号: 14370598
• 负责人: HATA Ryu-ichiro
• 金额: $ 8.38万
• 依托单位: Kanagawa Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14370598/
• 关键词: collagen; differentiation of osteoblasts; a long-acting vitamin C; 1α, 25 (OH)_2 vitamin D_3; human osteoblasts; MG63; 活性持続性ビタミンC

In order to investigate the mechanisms by which 1α25 (OH)_2 vitamin D_3 (VD_3) stimulates the differentiation of human osteoblasts, we cultured MG-63, which is a human osteoblastic cell line, in the presence or absence of VD_3 and/or L-ascorbic acid 2-phosphate (Asc 2-P), a long-acting vitamin C derivative. The cell growth rate was decreased by the presence of VD_3 in the culture medium. Type I collagen synthesis and alkaline phosphatase (ALP) activity, which are markers of early stage osteoblast differentiation, were stimulated by the presence of VD_3 as well as by that of Asc 2-P. The co-presence of Asc 2-P and VD_3 had a synergistic effect on the collagen synthesis and ALP activity of the cells. Inhibition of collagen synthesis by the addition of inhibitors of collagen synthesis to the medium attenuated the stimulative effect of VD_3 and Asc 2-P on the ALP activity. Transfection of the cells with siRNA-expressing vectors for COL1A1 decreased the expression level of ALP mRNA in addit … More ion to that of COL1A1. On the other hand, ALP activity was significantly increased, and the growth rate was decreased, when the cells were cultured on type I collagen-coated dishes. These effects were not seen when the cells were cultured on dishes coated with heat-denatured collagen. VD_3 also increased the mRNA levels for Runx2 and osterix, which are transcription factors critical for osteoblast differentiation, as well as those of differentiation markers such as bone/liver/kidney type ALP,COL1A1, (the gene for the α1 chain of type I collagen), and osteocalcin, in the cells. Normal human osteoblasts and human bone marrow-derived mesenchymal stem cells (hBMSC) showed quite similar responses to VD_3. These results indicate that VD_3 -stimulated gene expression of type I collagen and that mature type I collagen produced in the presence of Asc 2-P mediates at least a part of the stimulative effects of Asc 2-P and VD_3 on the differentiation of these human osteoblastic cells. Levels of mRNAs for ALP and COL1A1were increased, but the level of Runx2 was decreased, by the expression of osterix in MG-63 cells. These results also suggest that VD_3 controls the growth and differentiation of human osteoblastic cells by regulating the gene expression of osteoblast-related transcription factors as well as that of type I collagen, and that the co-presence of both signals is essential for VD_3 to express full activity toward the differentiation of human osteoblasts. Less

为了研究1α25（OH）_2维生素D_3（VD_3）刺激人类成骨细胞的分化的机制，我们培养了MG-63，它是人类成骨细胞系，在VD_3和/或l- cascorbic Acid的存在下，是人类成骨细胞系的分化。通过培养基中VD_3的存在降低了细胞生长速率。 I型胶原蛋白合成和合金线磷酸酶（ALP）活性是早期成骨细胞分化的标志物，通过VD_3的存在以及ASC 2-P的存在刺激。 ASC 2-P和VD_3的共呈现对细胞的胶原蛋白合成和ALP活性具有协同作用。通过在培养基中添加胶原蛋白合成的抑制剂来抑制胶原蛋白合成，从而减弱了VD_3和ASC 2-P对ALP活性的刺激作用。用表达siRNA的载体将细胞转染的COL1A1降低了ALP mRNA的表达水平，而col1a1则更多。另一方面，当细胞在I型胶原蛋白涂层的菜肴上培养时，ALP活性显着增加，并且生长速率降低。当将细胞培养在涂有热变性胶原蛋白的盘子上时，看不到这些影响。 VD_3还提高了Runx2和Osterix的mRNA水平，它们是成骨细胞分化至关重要的转录因子，以及分化标记的转录因子，例如骨/肝脏/肾脏型ALP，COL1A1，（I型I型胶原蛋白的α1基因）和细胞中的Osteocalcin。正常的人骨细胞和人骨骨髓衍生的间充质干细胞（HBMSC）对VD_3的反应非常相似。这些结果表明，I型胶原蛋白的VD_3刺激基因表达以及在ASC 2-P介导的存在下产生的成熟的I型胶原蛋白至少至少一部分ASC 2-P和VD_3对这些人类成骨细胞的分化的刺激作用。 ALP和COL1A1的mRNA水平升高，但通过MG-63细胞中的Osterix的表达，RUNX2的水平降低了。这些结果还表明，VD_3通过调节与成骨细胞相关的转录因子的基因表达以及I型胶原蛋白的基因表达来控制人类成骨细胞的生长和分化，并且这两种信号的共同点对于VD_3的共同呈现至关重要。较少的

项目成果

Kato, Y.: "Rho A⇒phospholipase⇒p38 MAP kinase signaling is an extracellular low PH responsible pathway fro matrix metalloproteinase-9 expression in mouse metastatic melanoma cells"Bull. Kanagawa Dent. Coll.. 30-1. 25-27 (2002)

Kato, Y.：“Rho A⇒磷脂酶⇒p38 MAP 激酶信号传导是小鼠转移性黑色素瘤细胞中基质金属蛋白酶 9 表达的细胞外低 PH 信号传导途径”，Kanakawa Dent. 25-27。 2002）

頭頸部癌抑制剤および医薬組成物

头颈癌抑制剂及药物组合物

• 发表时间: 2005

Izukuri K.et al.: "Development of cDNA microarray for analysis of gene expression profiles during the differentiation process in the craniomandibular system and for molecular diagnosis of craniomandibular disorders."Bull.Kanagawa Dent.Col.. 30(2). 137-140

Izukuri K.等人：“开发 cDNA 微阵列，用于分析颅颌系统分化过程中的基因表达谱以及颅颌疾病的分子诊断。”Bull.Kanakawa Dent.Col. 30(2)。

Kato Y., et al.: "Genetic Disruption of SPARC decreases in ERCCI expression and induces Skin Papilloma Formation"Bull.Kanagawa Dent.Col.31. 31. 71-73 (2003)

Kato Y.等人：“SPARC 的基因破坏降低了 ECCI 表达并诱导皮肤乳头状瘤形成”Bull.Kanakawa Dent.Col.31。

Extracellular Acidic pH and Hypoxia Regulate Expression of Metastasis-related Genes through Independent Mechanism.

细胞外酸性pH和缺氧通过独立机制调节转移相关基因的表达。

• 发表时间: 2004
• 期刊: Bull.Kanagawa Dent.Col. 32
• 作者: Momoi Y;Ariji Y;Katsumata A;Yoshida K;Masuoka N;Nawa H;Ariji E;Goto S;Maehata et al.;Kato Y.et al.;Kato Y.et al.;Kato Y.et al.;Kato Y.et al.;Kato Y.et al.;Schwarze U. et al.;Takamizawa S. et al.;Schwarze U.et al.;Takamizawa S.et al.;Baba Y.et al.;Schwarze U. et al.;Takamizawa S. et al.;Kato Y.et al.
• 通讯作者: Kato Y.et al.