南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (08): 1207-.

• • 上一篇    下一篇

LASP1下调和ferritin上调在rhBMP-2 诱导的比格犬BMSCs成骨分化中起重要作用

胡稷杰，刘亚伟，何敏毅，余斌，王钢   

  • 出版日期:2013-08-20 发布日期:2013-08-20

Recombinant human bone morphogenetic protein-2 down-regulates LASP1 and up-regulates ferritin during osteogenic differentiation of beagle dog bone marrow mesenchymal stem cells

  • Online:2013-08-20 Published:2013-08-20

PDF

1106

摘要： 目的建立稳定的比格犬骨髓间充质干细胞（BMSCs）诱导成骨分化模型；鉴定参与比格犬BMSCs细胞成骨分化过调控的
蛋白质，探讨其可能的调控机制。方法分离培养比格犬BMSCs细胞，rhBMP-2诱导细胞成骨分化7 d，基于蛋白质双向凝胶电
泳的蛋白质组学分析成骨分化组与对照组差异表达的蛋白质，对感兴趣的蛋白质LASP1、ferritin 轻链和重链表达情况用
Q-PCR、Western blot 进行验证。结果成功诱导比格犬BMSCs成骨分化；蛋白质组学分析获得rhBMP-2 诱导上调的蛋白质9
个，下调的蛋白质11 个。荧光定量PCR技术和Western blot 对LASP1 和ferritin 表达情况进行了验证，发现rhBMP-2 诱导7 d
后，LASP1 显著下调而ferritin 显著上调，与蛋白质组学结果一致。结论LASP1 在调节细胞骨架活性方面发挥重要功能，而
ferritin 是细胞铁离子稳态维持的重要分子，提示这两种蛋白质在rhBMP-2 诱导的比格犬BMSCs成骨分化过程中发挥重要
作用。

Abstract: Objective To identify the proteins involved in osteogenic differentiation of beagle dog bone marrow mesenchymal
stem cells (BMSCs) and explore its possible regulation mechanism of osteogenic differentiation of BMSCs. Methods Cultured
beagle dog BMSCs were induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) for 7 days. The
differentially expressed proteins between cells with osteogenic differentiation and control cells were identified by proteomics
analysis based on two-dimensional gel electrophoresis. Q-PCR and Western blotting were used to verify the interested protein
of LASP1, ferritin light chain and heavy chain. Results Osteogenic differentiation was induced successfully in the BMSCs.
Twenty differentially expressed proteins were identified by proteomic analysis, including 9 down-regulated and 11
up-regulated ones. Q-PCR and Western blotting demonstrated a significant reduction of LASP1 expression and significant
up-regulation of ferritin in the BMSCs after a 7-day induction with rhBMP-2. Conclusion LASP1, which plays an important
role in the regulation of the activity of the cytoskeleton, and ferritin, an important molecule in cellular iron homeostasis, can be
critical in the osteogenic differentiation of beagle dog BMSCs induced by rhBMP-2.