Differential proteome analysis of replicative senescence in rat embryo fibroblasts
Differential proteome analysis of replicative senescence in rat embryo fibroblasts
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DOI：
10.1074/mcp.m100028-mcp200
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发表时间：
2002-04-01
影响因子：
7
通讯作者：
Jat, PS
中科院分区：
文献类型：
--
作者：
Benvenuti, S;Cramer, R;Jat, PS
正常的体细胞经历有限数量的分裂，然后停止分裂，而癌细胞能够无限期地增殖。为了确定限制有丝分裂潜力的潜在机制，进行了连续传代大鼠胚胎成纤维细胞复制性衰老的二维差异蛋白质组分析。运行、策划和分析了各自含有超过1200个斑点的不依赖于三个周期的二维凝胶。这揭示了49个点，其表达改变超过2倍。其中，42个点产生了积极的蛋白质鉴定质谱包括各种细胞骨架，热休克和代谢蛋白，以及蛋白质参与运输，分化，蛋白质合成，营业额，和修改。这些包括凝溶胶蛋白，一种乳腺癌的候选肿瘤抑制因子，和α-葡萄糖苷酶11，一种包括klotho的葡萄糖苷酶家族的成员;小鼠中klotho表达的缺陷导致类似人类衰老的综合征。还鉴定了TUC-1、-2、-4和-4 β（TUC家族成员，对神经元分化至关重要）表达的变化。一些确定的变化也显示发生在另外两种衰老模型中，即REF 52细胞的早衰和小鼠胚胎成纤维细胞的复制性衰老。这些候选蛋白中的大多数在复制性衰老中以前未被识别。他们现在扮演着一个新的角色。
Normal somatic cells undergo a finite number of divisions and then cease dividing whereas cancer cells are able to proliferate indefinitely. To identify the underlying mechanisms that limit the mitotic potential, a two-dimensional differential proteome analysis of replicative senescence in serially passaged rat embryo fibroblasts was undertaken. Triplicate independent two-dimensional gels containing over 1200 spots each were run, curated, and analyzed. This revealed 49 spots whose expression was altered more than 2-fold. Of these, 42 spots yielded positive protein identification by mass spectrometry comprising a variety of cytoskeletal, heat shock, and metabolic proteins, as well as proteins involved in trafficking, differentiation, and protein synthesis, turnover, and modification. These included gelsolin, a candidate tumor suppressor for breast cancer, and alpha-glucosidase 11, a member of the family of glucosidases that includes klotho; a defect in klotho expression in mice results in a syndrome that resembles human aging. Changes in expression of TUC-1, -2, -4, and -4beta, members of the TUC family critical for neuronal differentiation, were also identified. Some of the identified changes were also shown to occur in two other models of senescence, premature senescence of REF52 cells and replicative senescence of mouse embryo fibroblasts. The majority of these candidate proteins were unrecognized previously in replicative senescence. They are now implicated in a new role.