The Key Laboratory of Education Ministry for Cell Biology and Tumor Cell Engineering, School of Life Science, Xiamen University, Xiamen, China.
Anat Rec (Hoboken). 2010 May;293(5):813-20. doi: 10.1002/ar.21074.
The objective of this study was to investigate altered expressions of nuclear matrix proteins (NMPs) of human osteosarcoma (OS) MG-63 cells during curcumin-induced apoptosis of human OS MG-63 cells. MG-63 cells were cultured with curcumin (7.5 mg/L) for 72 hr. Morphological alterations of cells were captured using light microscopy and transmission electron microscopy, and cell cycle distribution was estimated by flow cytometry. NMPs were selectively extracted and subjected to two-dimensional gel electrophoresis (2-DE) analysis. Western blots were performed to determine changes in the expression levels of specific NMPs. The results demonstrated that typical characteristics of apoptosis were observed. Cellular chromatin agglutinated, cell nuclei condensed, and apoptotic bodies were formed after treatment with curcumin. The 2-DE results displayed 27 NMPs, 21 of which were identified to have change in expression levels significantly during apoptosis. The altered expressions of three of these NMPs (nucleophosmin, prohibitin, and vimentin) were further confirmed by immunoblotting. These findings indicated that the apoptosis of MG-63 cells was accompanied by the expression alteration of NMPs. Our results might help to reveal the relationship between NMPs and the regulation of gene expression in the process of apoptosis, as well as provide the basic concepts for future studies on the mechanisms of apoptosis and the therapy for bone diseases.
本研究旨在探讨姜黄素诱导人骨肉瘤(OS)MG-63 细胞凋亡过程中核基质蛋白(NMPs)的表达变化。将 MG-63 细胞用姜黄素(7.5mg/L)培养 72 小时。用光镜和透射电镜观察细胞形态变化,用流式细胞术估计细胞周期分布。选择性提取 NMPs 并进行二维凝胶电泳(2-DE)分析。Western blot 用于测定特定 NMPs 表达水平的变化。结果表明,观察到典型的凋亡特征。用姜黄素处理后,细胞染色质聚集,细胞核浓缩,形成凋亡小体。2-DE 结果显示 27 种 NMPs,其中 21 种在凋亡过程中表达水平明显变化。这三种 NMPs(核仁磷酸蛋白、抑制素和波形蛋白)的改变表达进一步通过免疫印迹得到证实。这些发现表明 MG-63 细胞的凋亡伴随着 NMPs 的表达改变。我们的结果可能有助于揭示 NMPs 与凋亡过程中基因表达调控之间的关系,并为凋亡机制和骨疾病治疗的未来研究提供基础概念。