抑制性消减杂交筛选细胞衰老相关基因

Identifying cellular senescence-associated genes by suppression subtractive hybridization

目的 研究人胚肺二倍体成纤维细胞 2BS在衰老过程中的基因表达变化。 方法 对年轻 2BS细胞〔低于 30PD(populationdoubling)〕和衰老 2BS细胞 (高于 5 5PD)样品进行双向抑制性消减杂交 ,并筛选分析。 结果 构建了 2个抑制性消减文库 ,分别代表年轻细胞和衰老细胞高表达基因 ,点杂交筛选获得 30个差异表达片断 (差异比例大于 2 )包含多种细胞功能的变化。部分差异基因表达在新生儿脐带血和老年人外周血样中差异也有显著性。 结论 首次报道了RBM4、FBXO7、TOM1等基因在 2BS细胞衰老时的表达变化 ,并表明体外培养细胞与个体的衰老变化过程有一定相似性。

Objective The differentially expressed genes in the progress of cellular senescence were studied. Methods Suppression subtractive hybridization was used to young and senescent human diploid fibroblast 2BS cells, then screening and analyzing the subtractive libraries. Results Two kinds of subtraction libraries were obtained, respectively representing the differential genes with higher expression in young cells or senescent cells. Screened by dot blot, thirty of genes with differential ratio larger than 2 were sequenced, showing changes in much of cell functions. Some of them also had expression change between blood samples from newborn or old people. Conclusions The differential expression of RBM4?FBXO7?TOM1 is reported during 2BS cell senescence for the first time. There are the similarities during the senescence of culture cell and organism.