摘要
目的应用蛋白质组学方法分析人成骨样细胞Saos-2在牵张应力作用下蛋白质表达的差异,为更全面地阐明成骨细胞对牵张应力的反应机制提供分子基础。方法将Saos-2分为加力组与对照组,采用Flexcell牵张应力加载系统,用12%大小的应力值进行力学刺激24 h后,应用蛋白质双向凝胶电泳分离蛋白样品,质谱技术鉴定差异表达蛋白点,并用生物信息学分析差异蛋白参与的生物学过程和主要功能。结果对照组和加力组分别得到(1031±41)和(928±25)个蛋白质点,质谱鉴定出肽酰脯氨酰异构酶A样3、线粒体ATP合成酶、抗氧化蛋白1、丝切蛋白1、蛋白磷酸酶1、延伸因子2等17个表达增高的差异蛋白质,涉及应激反应、能量代谢、细胞增殖、细胞骨架重建、信号转导及成骨分化等生物学功能。结论成骨细胞在机械牵张应力作用下蛋白表达发生显著变化,这些差异蛋白参与了成骨细胞力学反应机制的不同过程。
Objective To identify the differentially expressed proteins and clarify the major proteins involved in the molecular mechanism of osteoblasts under mechanical strain loading.Method Saos-2 osteoblastic cells were subjected to 12% elongation for 24 hours by using Flexcell strain loading system.Proteins extracted from Saos-2 cells were separated by two-dimensional electrophoresis（2-DE）.Differential expressed protein spots among groups were submitted to matrix-assisted laser desorption/ionization time of flight mass spectrometer（MALDI-TOF MS） assay and peptide mass fingerprinting（PMF） identification.The Swiss-Prot and NCBI databases were used to obtain further information about proteins identified.Results Saos-2 stimulated by mechanical strain showed a significant difference in 2-DE system compared with the control group.A total of（1031 ±41） or（928 ± 25） protein spots were resolved by 2-DE of controls or experimental groups extractions respectively.17 significant up-regulated proteins were identified.These associated proteins fell into 6 groups,including stress reaction,en-ergy metabolism,cell proliferation,reconstruction of cytoskeleton,signaling and osteogenesis.Conclusions The Saos-2 can express differential proteins stimulated by mechanical strains and these proteins may play an im-portant role in molecular mechanism of osteoblasts under mechanical strain loading.
出处
《医用生物力学》
EI
CAS
CSCD
2010年第6期406-411,共6页
Journal of Medical Biomechanics
基金
国家自然科学基金资助项目(30870594)
关键词
机械牵张应力
成骨细胞
蛋白质组学
细胞增殖
信号转导
Mechanical strain
Osteoblastic cells
Proteomics
Cell proliferation
Signal transduction
