Expression of Pcp4 gene during osteogenic differentiation of bone marrow mesenchymal stem cells in vitro
Expression of Pcp4 gene during osteogenic differentiation of bone marrow mesenchymal stem cells in vitro
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DOI：
10.1007/s11010-007-9652-x
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发表时间：
2008-02-01
影响因子：
4.3
通讯作者：
Liu, Lei
中科院分区：
文献类型：
--
作者：
Xiao, Jingang;Wu, Yao;Liu, Lei
本研究建立了大鼠骨髓间充质干细胞（BMSCs）体外成骨诱导分化模型，探讨BMSCs成骨的机制及相关基因功能。采用含β-甘油醛-3-磷酸、L-抗坏血酸、地塞米松和1，25 -2（OH）（2）维生素D-3的α-最小必需培养基诱导第三代BMSCs向成骨细胞分化，然后应用基因芯片技术（也称为微阵列技术）进行基因表达的整体筛选。采用实时定量PCR（Real-time PCR）检测骨髓基质干细胞成骨分化过程中调控基因mRNA表达的时间变化。利用生物信息学分析来确定所鉴定的成骨相关基因的功能意义。通过基因芯片筛选，浦肯野细胞蛋白4（Pcp 4）mRNA表达被鉴定为在BMSCs成骨分化过程中上调。实时荧光定量PCR分析证实，Pcp 4 mRNA的表达增加，在成骨分化的BMSCs的表达呈上升趋势，在第14天达到高峰。生物信息学分析确定Pcp 4为参与钙沉积和钙调蛋白依赖性蛋白激酶调节的基因。因此，我们假设Pcp 4诱导的钙沉积形成矿物质结节和某些信号转导通路的BMP的调制介导的BMSCs的成骨分化的Pcp 4。
In this study, we established an in vitro model of osteogenic-inductive differentiation of rat bone marrow mesenchymal stem cells (BMSCs) to determine the mechanisms and relative gene function underlying BMSCs osteogenesis. Osteoplastic differentiation of the third generation BMSCs was induced with the alpha-minimal essential medium containing beta-glyceraldehyde-3-phosphate, L-ascorbic acid, dexamethasone and 1,25-2(OH)(2) vitamin D-3 prior to applying gene chip technology (also called microarray technology) for global gene expression screening. Real-time quantitative PCR (Real-time PCR) was used to determine the temporal profile of mRNA expression of regulated genes during osteogenic differentiation of BMSCs. A bioinformatic analysis was utilized to determine the functional significance of the identified osteogenic-related genes. Purkinje cell protein 4 (Pcp4) mRNA expression was identified by the gene chip screening as being up-regulated during osteoplastic differentiation of BMSCs. Real-time PCR analysis confirmed the increased expression of Pcp4 mRNA expression during osteoplastic differentiation of BMSCs with an upward trend that peaked at day 14. The bioinformatic analysis identified Pcp4 as a gene involved in the deposition of calcium and the modulation of CaM-dependent protein kinase. Thus, we hypothesize that Pcp4 osteoplastic differentiation of BMSCs is mediated in part via Pcp4-induced calcium deposition to form mineral nodules and modulation of certain signal transduction pathways of BMPs.