Cellular signal transduction system in prostaglandin-stimulated osteoblast-like MC3T3-E1 cells : Phospholipid turnover and protein tyrosine phosphorylation

前列腺素刺激的成骨细胞样 MC3T3-E1 细胞中的细胞信号转导系统：磷脂周转和蛋白质酪氨酸磷酸化

• 批准号: 06672000
• 负责人: NAKASHIMA Shigeru
• 金额: $ 1.41万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06672000/
• 关键词: osteoblast cells; prostaglandins; phospholipase C; phospholipase D; protein kinase C; protein tyrosine kinase; プロスタグランシン; プロスタグランジンFSα; ジアシルグリセロール; ホスファチジン酸; Cキナーゼ; MAPキナーゼ

The combination of beta-glycerophosphate and ascorbic acid increased alkaline phosphate (ALP) activity, a maraker for osteoblastic differentiation, in clonal osteoblast like cell line MC3T3-E1. The increase was detected as 24 hours after the addition of these agents to growth medium and reached almost maximal level at 5 days of culture. Although increase in intracellular cyclic AMP level is known to induce osteoblastic acell differentiation, these treatment did not cause significant increase in cyclic AMP.Treatment of beta-glycerophsphate and ascorbic acid suppressed activaaton of phosphoinositide-specific phospholipase C (PI-PLC) and phospholipase D (PLD) in response to prostaglandin F_<2alpha> (PGF_<2alpha>) by 20-30% and 15%, respectively. Western blot analyzes with subtype-specific PI-PLC antibodies revealed that in cells treated for 5 days with these differentiation inducers, the levels of PI-PLCbeta1 and PI-PLCbeta3 decreased to 37% and 67% compared to those of undifferentiated control cells, respectively. However, the level of PLC_<gamma>1 and those of protein kinase C isozymes (alpha, beta1, beta2, delta, epsilon, rheta, zeta) remained unaltered. PGF_<2alpha> also stimulated protein tyrosine phosphorilations and activation of mitogen-activated protein (MAP) kinases. These responses were not affected by differentiation induceres. These results suggest that the suppressed expression of PI-PLCbeta1 and PI-PLCbeta3 are closely related to osteoblast cell differentiation.

联合应用β-甘油磷酸和抗坏血酸可增加成骨细胞克隆细胞系MC3T3-E1中碱性磷酸酶(ALP)的活性，碱性磷酸酶是成骨细胞分化的标志物。在加入这些药剂后24小时检测到细胞增殖，在培养5天时几乎达到最高水平。虽然细胞内环磷酸腺苷水平的升高可以诱导成骨细胞分化，但这些处理并没有引起环磷酸腺苷的显著增加。β-甘油磷酸和抗坏血酸的处理分别抑制了前列腺素F_&lt；2pha&gt；(PGF_&lt；2pha&gt；)对磷脂酶C(PI-PLC)和磷脂酶D(PLD)的激活20-30%和15%。用亚型特异性PI-PLC抗体进行Western印迹分析表明，在这些分化诱导剂处理5d的细胞中，PI-PLCbeta1和PI-PLCbeta3的表达水平分别下降到未分化对照细胞的37%和67%。然而，PLC_1和蛋白激酶C同工酶(α、β1、β2、Delta、epsilon、Rheta、Zeta)的水平没有变化。PGF还可刺激蛋白酪氨酸的磷酸化和丝裂原活化蛋白(MAP)的激活。这些反应不受分化诱导剂的影响。这些结果提示PI-PLCbeta1和PI-PLCbeta3的抑制表达与成骨细胞的分化密切相关。

项目成果

Kato, Y., Banno, Y., Nakashima, S.and Oka, N.: "Effect of prostaglandin F2a on osteoblast-like cell MC-3T3-E1 in the early stage of differentiation" J.Jpn.Stomatol.Soc.45(1). 1-12 (1996)

Kato, Y.、Banno, Y.、Nakashima, S. 和 Oka, N.：“前列腺素 F2a 对分化早期的成骨细胞样细胞 MC-3T3-E1 的影响”J.Jpn.Stomatol.Soc。

Sugiyama T.et al.: "Prostaglandin F2d-stimulated Phospholipase D activation in osteo blast-like Mc3T3-El cells:Involvement in sustained 1.2-diacylgbjcerol production" Biochemical Journal. 298. 479-484 (1994)

Sugiyama T.等人：“成骨细胞样 Mc3T3-El 细胞中前列腺素 F2d 刺激的磷脂酶 D 激活：参与持续的 1.2-二酰基甘油生产”生物化学杂志。

加藤幸弘: "骨芽細胞様細胞MC3T3E1の分化初期応答におけるプロスタグランジンF_<2α>の作用" 日本口腔外科学雑誌. 45. 1-12 (1996)

Yukihiro Kato：“前列腺素F_<2α>对成骨细胞样细胞MC3T3E1的早期分化反应的影响”日本口腔外科杂志45. 1-12 (1996)。

Sugiyama,T.: "Prostaglandin F2 α-stimulated phosphlipase D activation in osteoblast-like MC-3T3-E1 cells." Biochem.J.298. 479-484 (1994)

Sugiyama, T.：“前列腺素 F2 α 刺激的成骨细胞样 MC-3T3-E1 细胞中的磷脂酶 D 激活。Biochem.J.298（1994）。”

加藤 幸弘 他: "骨芽細胞様細胞MC3T3-E1の分化初期応答におけるPGF2αの作用" 日本口腔外科学会雑誌. 45. 1-12 (1996)

Yukihiro Kato 等：“PGF2α 对成骨细胞样细胞 MC3T3-E1 的早期分化反应的影响”日本口腔颌面外科学会杂志 45. 1-12 (1996)。