Genome-wide pathways analysis of nickel ion-induced differential genes expression in fibroblasts
Genome-wide pathways analysis of nickel ion-induced differential genes expression in fibroblasts
复制标题
镍离子诱导的成纤维细胞差异基因表达的全基因组通路分析
DOI：
10.1016/j.biomaterials.2009.12.044
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发表时间：
2010-03-01
期刊：
影响因子：
14
通讯作者：
Lu, Zuhong
中科院分区：
文献类型：
--
作者：
Lue, Xiaoying;Lu, Huiqin;Lu, Zuhong
为了揭示Ni2+与细胞相互作用的分子机制，利用基因芯片技术和基因芯片技术，研究了100MMNi2+作用于小鼠成纤维细胞(L 92 9)12、2 4、48和72 h后基因表达谱的变化，确定了重要的生物学途径。芯片数据通过实时荧光定量聚合酶链式反应进行验证。采用四甲基偶氮唑蓝比色法和流式细胞仪检测细胞对Ni2+的反应。结果发现，Ni2+影响了6条主要的生物学途径，涉及118个差异表达基因。进一步的分析表明，细胞暴露于Ni2+可能通过调节低氧诱导基因的表达而引起细胞对低氧的一系列反应，并导致不可逆的DNA损伤。细胞周期分析结果显示，Ni2+能抑制S期细胞的复制，这与流式细胞仪实验结果一致。与以往基于常规分子生物学实验的研究相比，本工作不仅间接验证了其他研究小组的发现，而且还获得了与细胞-Ni2+相互作用相关的几个发现，如抑制电子传递链和细胞外基质(ECM)胶原的积累。本研究的常规方法不仅可以分析基因表达谱，而且可能为解释细胞-生物材料相互作用的分子机制提供一种更方便、更有效的方法。(C)2009爱思唯尔有限公司。保留所有权利。
To reveal molecular mechanisms of the interaction between Ni2+ and cells, cDNA microarray technology and GenMAPP analysis were utilized to investigate changes of gene expression profile and identify significant biological pathways in mouse fibroblast cells (L-929) treated by 100 mu m Ni2+, for 12, 24, 48 and 72 h, respectively. The microarray data was validated by real-time PCR. Methylthiazolte-trazolium (MTT) analysis and flow cytometry experiment were used to assess the cellular response of L-929 cells to Ni2+. It was found that six main biological pathways were affected by Ni2+, with 118 differentially expressed genes involved. Further analysis illuminated that the exposure of cells to Ni2+ may evoke series of cellular responses to hypoxia by regulating hypoxia-inducible gene expression and cause irreversible DNA damage. Cell cycle pathway analysis results showed DNA replication in S phase could be inhibited by Ni2+, which was consistent with the data gained from flow cytometry experiment. Compared to previous researches based on conventional molecular biology experiments, the present work has not only indirectly validated the findings of other groups but also obtained several discoveries related to cell-Ni2+ interaction, such as inhibition of electron transport chain and accumulation of extracellular matrix (ECM) collagens. The routine of the present study not only can analyze gene expression profile but also may provide a more convenient and efficient approach to explain molecular mechanisms of cell-biomaterial interaction. (C) 2009 Elsevier Ltd. All rights reserved.