国家重点基础研究发展规划资助项目(G2000517001和G1999053906)和国家自然科学基金重点资助项目(39930170).
This work was supported by grants from The Special Funds for Major State Basic Research of China (G2000517001 and G1999053906), The National Natural Sciences Foundation of China (39930170).
以50 μmol/L H2O2作用体外培养的人胚肺二倍体成纤维细胞4次,使之出现不可逆的衰老表型.提取年轻细胞及H2O2处理早老细胞的mRNA,以荧光物Cy3标记年轻细胞cDNA,Cy5标记H2O2处理的细胞cDNA,并与点有4 096条人类基因的芯片杂交,利用计算机数据处理判断基因是否存在表达差异.结果显示:有123种基因的表达变化较显著,这些基因参与细胞周期进程、细胞代谢及蛋白质修饰、细胞外基质及细胞骨架蛋白的形成和调节、炎症反应、调节受体酪氨酸蛋白激酶和G蛋白耦联受体信号转导.
The senescent phenotype of human embryo lung diploid fibroblasts was obtained by four treatments of early passage cells with 50 μmol/L H2O2 for 30 min every 2 days. After total mRNA extraction from the two kinds of cells, the young cells and H2O2-induced premature senesence cells,the cDNA of two kinds of cells were labled by Cy3 and Cy5 respectively were hybridized with microarray containing 4 096 human genes. The gene expression changes were identified by GnePix 4000B and GenePix 3.0. 123 genes changed their expression significantly, which involved in progress of cell cycle,metabolism and protein processing, formation and modification of cytoskeleton and extracellular matrix and signal transduction, and the most interesting finding was that secretory function seemed to be enhanced in premature senescence of fibroblasts.
马宏,张宗玉,童坦君. H2O2诱发人成纤维细胞衰老样变化的基因表达谱[J].生物化学与生物物理进展,2003,30(1):72-77
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