Role of basic helix-loop-helix SCL protein in osteoclast differentiation

碱性螺旋-环-螺旋SCL蛋白在破骨细胞分化中的作用

• 批准号: 07671984
• 负责人: AMANO Shigeru
• 金额: $ 1.41万
• 依托单位: Meikai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671984/
• 关键词: Osteoclast; SCL; Bone-resorbing function; Transcriptional factor; 分化

The present study was conducted to determine whether SCL (stem-cell leukemia) protein, a member of basic helix-loop-helix (bHLH) class of transcriptional factors, implicates in osteoclast formation or function of mature osteoclasts.Antisense s-oligonucleotide (2muM) includeing the translation initiation region of the mouse SCL gene inhibites pit formation in a dose-dependent manner, but not inhibites TRAP-positive cell formation. The SCL gene expression was observed by using the RT-PCR technique. Although the constitutive SCL gene expression decreases in time-dependent manner when 14-day-old mouse embryonic calvarial cells are cultured in the absence of 1alpha, 25- (OH)_2D_3, in the presence of 1alpha, 25- (OH)_2D_3, the SCL gene expression decreases transiently on day 3 after the initiation of the culture, and then the gene expression increases on day 5 again. In addition, the SCL gene expression is also observed in mature osteoclast-enriched population. In constrast with these observation, the constitutive expression of SCL gene in early myeloid cell line M1 cells was disappeared in 1alpha, 25- (OH)_<>D_3 treatment-time dependent manner. Antisense SCL s-oligonucleotide inhibits pit formation evenwhen the antisense oligomer is added to TRAP-positive cell forming the late stage cells. In addition, the inhibition of pit formation by antisense SCL s-oligonucleotide is observed in bone cells including mature osteoclasts prepared from 6-day-old mice femora. These results suggest that SCL protein is involved in the exhibition of bone-resorbing function of osteolasts.

本研究旨在确定 SCL（干细胞白血病）蛋白（碱性螺旋-环-螺旋 (bHLH) 类转录因子的成员）是否与破骨细胞的形成或成熟破骨细胞的功能有关。反义 s-寡核苷酸 (2μM ) 包括小鼠 SCL 基因的翻译起始区，以剂量依赖性方式抑制凹陷形成，但不抑制 TRAP 阳性细胞形成。采用RT-PCR技术观察SCL基因表达。尽管在不存在 1α, 25- (OH)_2D_3 的情况下培养 14 日龄小鼠胚胎颅骨细胞时，在存在 1α, 25- (OH)_2D_3 的情况下，组成型 SCL 基因表达会以时间依赖性方式降低， SCL基因表达在培养开始后第3天短暂下降，然后在第5天基因表达再次增加。此外，在成熟的破骨细胞丰富的群体中也观察到了SCL基因的表达。与这些观察相反，早期骨髓细胞系M1细胞中SCL基因的组成型表达以1α，25-(OH)_<>D_3治疗时间依赖性方式消失。即使当反义寡聚物被添加到形成晚期细胞的TRAP阳性细胞时，反义SCL s-寡核苷酸也抑制凹坑形成。此外，在从 6 日龄小鼠股骨制备的骨细胞（包括成熟破骨细胞）中观察到反义 SCL s-寡核苷酸对凹坑形成的抑制。这些结果表明SCL蛋白参与成骨细胞骨吸收功能的发挥。

项目成果

Hanazawa,S.: "Porphyromonas gingivalis fimbria-stimulated bone resorption in vitro is inhibited by a tyrosine kinase inhibitor." Infect.Immun.63. 2374-2377 (1995)

Hanazawa,S.：“酪氨酸激酶抑制剂可抑制体外牙龈卟啉单胞菌菌毛刺激的骨吸收。”

Amano,S.: "Prostaglandin E_2stimulates osteoclast formation via endogeneous IL-1β expressed through protein kinase A." J.Immunol.156. 1931-1936 (1996)

Amano, S.：“前列腺素 E_2 通过蛋白激酶 A 表达的内源性 IL-1β 刺激破骨细胞形成。J.Immunol.1931-1936 (1996)。

Amano, S.: "Prostaglandin E_2 stimulates osteoclast formation via endogeneous IL-1beta expressed through protein kinase A." J.Immunol.Vol.156. 1931-1936 (1996)

Amano, S.：“前列腺素 E_2 通过蛋白激酶 A 表达的内源性 IL-1β 刺激破骨细胞形成。”

Shigeur Amano: "Prostaglandin E_2 Stimulates Osteoclast Formation via Endogenous Interleukin-1β Expressed Through Protein Kinase A." J. Immunol.156. (1996)

Shigeur Amano：“前列腺素 E_2 通过蛋白激酶 A 表达的内源性白细胞介素 1β 刺激破骨细胞形成。J.Immunol.156。”

Hanazawa,S.: "Prophyromonas gingivalis fimbria-stimulated bone resorption in vitro is inhibited by a tyrosine kinase inhibitor." Infect.Immun.63・6. 2374-2377 (1995)

Hanazawa, S.：“酪氨酸激酶抑制剂可抑制牙龈卟啉单胞菌菌毛刺激的骨吸收。”Infect.Immun.63·6 (1995)。