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通过E2F-1过表达转化的已建立细胞系含有野生型p53。

Established cell lines transformed by E2F-1 overexpression contain wild-type p53.

作者信息

Sladek T L

机构信息

Department of Microbiology and Immunology, Finch University of Health Sciences, Chicago Medical School, Illinois 60064-3095, USA.

出版信息

Cell Prolif. 1996 Nov;29(11):579-88. doi: 10.1111/j.1365-2184.1996.tb00973.x.

Abstract

Overexpression of genes encoding E2F transcription factors can transform some cultured cell lines and cause apoptosis of others. Apoptosis due to E2F overexpression requires the presence of wild-type p53. Cell lines in which stable E2F overexpression is possible might be expected, therefore, to contain mutant p53. In this report, it was asked whether endogenous p53 was mutant or wild-type in four established fibroblast cell lines that this laboratory previously showed stably overexpressed and were transformed by exogenous E2F-1. Unexpectedly, it was found that the p53 in these cells was wild-type by the criteria of immunoprecipitation with conformation-specific, p53 monoclonal antibodies and by transactivation of a p53-dependent reporter gene construct in transient transfection assays. These data indicate that stable overexpression of E2F-1 is possible in the presence of wild-type p53 and may result in cell transformation.

摘要

编码E2F转录因子的基因过表达可使一些培养的细胞系发生转化,而使另一些细胞系发生凋亡。E2F过表达引起的凋亡需要野生型p53的存在。因此,那些能够实现稳定E2F过表达的细胞系可能含有突变型p53。在本报告中,我们研究了本实验室先前发现的四种已建立的成纤维细胞系中内源性p53是突变型还是野生型,这些细胞系先前显示能稳定过表达并被外源性E2F-1转化。出乎意料的是,通过使用构象特异性p53单克隆抗体进行免疫沉淀以及在瞬时转染实验中对p53依赖性报告基因构建体的反式激活标准,发现这些细胞中的p53是野生型。这些数据表明,在野生型p53存在的情况下,E2F-1的稳定过表达是可能的,并且可能导致细胞转化。

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