Interaction of cytokines and cell adhesion molecules during osteoclastogenesis

破骨细胞生成过程中细胞因子和细胞粘附分子的相互作用

• 批准号: 11671089
• 负责人: INOUE Masaru
• 金额: $ 0.77万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671089/
• 关键词: osteoclast precursors; signal transduction; tyrosine phosphorylation; integrin; M-CSF; cell adhesion; 信号伝達系; 細胞外マトリックス; 破骨細胞; 細胞接着因子; マクロファージコロニー刺激因子

Most type of cells need to contact with extracellular matrix, which are mainly mediated with integrins. The signal transduction pathway through integrin shares many parts with that of cyokines. The expression of integrin avβ5 is abundant on osteoclast precursors, which need many kinds of cyokines, including M-CSF, for their maturation and survival. Therefore we explored interaction of integrin and cytokine signal pathway on osteoclast precursors.Osteoclast precursors were isolated from C3H mouse hindlimb bones and cultured in PFA dishes for 3 days with M-CSF. Osteoclast precursors were collected and cultured on plastic(A) or PFD(NA) dishes with(+) or without(-) M-CSF. Cell lysate from each group were fractionated on SDS-PAGE, blotted onto membrane and proved by anti-phosphorlated tyrosine.The 50kDa band were regulated by cell attachment and decreased in NA compared with A. The effect of M-CSF was not clear between NA- and NA+, the intensity of 50kDa band in A+ showed 1.3fold increase of that in A-.The 50kDa protein is the candidate of signal transduction molecules. Cell adhesion is necessary for the effect of M-CSF since the intensity of 50kDa band was up-regulated by M-CSF in A.

大多数类型的细胞需要与细胞外基质接触，而细胞外基质主要是由整合素介导的。通过整合素的信号转导途径与细胞因子的信号转导途径有许多共同之处。整合素avβ5在破骨细胞前体上表达丰富，而破骨细胞的成熟和存活需要多种细胞因子，包括M-CSF。因此，我们探索整合素和细胞因子信号通路在破骨细胞前体中的相互作用。从C3H小鼠后肢骨中分离破骨细胞前体，用M-CSF在PFA培养皿中培养3天。收集破骨细胞前体，在塑料(A)或PFD（NA）培养皿中培养（+）或不（-）M-CSF。各组细胞裂解液在SDS-PAGE上分离，印迹于膜上，用抗磷酸化酪氨酸进行鉴定。与A相比，M-CSF在NA-和NA+中的作用不明显，在A+中的50kDa条带强度比在A-中增加1.3倍。50kDa蛋白是信号转导的候选分子。细胞粘附是M-CSF发挥作用的必要条件，因为M-CSF在A中上调了50kDa波段的强度。

项目成果

Y. Seino, T, Moriwake, H. Tanaka, M. Inoue, S. Kanzaki, T. Tanaka, N. Matsuo, H. Niimi: "Molecular defects in achondroplasia and the effects of growth hormone treatment"Acta Paediatrica Suppl.. 428. 118-120 (1999)

Y. Seino, T, Moriwake, H. Tanaka, M. Inoue, S. Kanzaki, T. Tanaka, N. Matsuo, H. Niimi：“软骨发育不全的分子缺陷和生长激素治疗的影响”学报儿科增刊。

M. Inoue, H. Tanaka, T. Moriwake, M. Oka, C. Sekiguchi, Y. Seino: "Altered Biochemical Markers of Bone Turnover in Humans During 120 Days of Bed Rest."Bone. 26. 281-286 (2000)

M. Inoue、H. Tanaka、T. Moriwake、M. Oka、C. Sekiguchi、Y. Seino：“卧床休息 120 天期间人类骨转换的生化标志物发生改变”。

Y.Seino, et al.: "Molecular defects in achondroplasia and the effects of Growth hormone treatment"Acta Pediatrica Suppi.. 428. 118-120 (1999)

Y.Seino 等：“软骨发育不全的分子缺陷和生长激素治疗的影响”Acta Pediatrica Suppi.. 428. 118-120 (1999)

T.Miyamura, et al.: "The Effects of Bone Marrow Transplantation on X-linked Hypophosphatemic Mice"J. of Bone and Mineral Research. 15. 1451-1458 (2000)

T.Miyamura 等人：“骨髓移植对 X 连锁低磷血症小鼠的影响”J。

M.Inoue, et al.: "Altered Biochemical Markers of Bone Turnover in Human During 120 Days of Bed Rest"Bone. 26. 281-286 (2000)

M.Inoue 等人：“卧床休息 120 天期间人体骨转换的生化标志物发生改变”