Immortalization of human fibroblasts transformed by origin-defective simian virus 40.
Immortalization of human fibroblasts transformed by origin-defective simian virus 40.
复制标题
起源缺陷型猿猴病毒 40 转化的人成纤维细胞的永生化。
DOI：
10.1128/mcb.7.8.2794-2802.1987
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发表时间：
1987
影响因子：
5.3
通讯作者：
Ozer,HL
中科院分区：
文献类型：
--
作者：
Neufeld,DS;Ripley,S;Henderson,A;Ozer,HL
猴病毒40(SV40)介导的人二倍体成纤维细胞转化为研究细胞培养中的“衰老”和癌变提供了有效的实验系统。然而，以前的解释可能因为半允许的病毒与细胞的相互作用而变得复杂。在早期的研究中，我们已经证明SV40的复制缺陷突变体的DNA可以有效地转化人二倍体成纤维细胞系HS74，该突变体含有用于DNA合成的病毒起源缺失(SVori-)。在目前的研究中，我们发现，与HS74或包含完整病毒基因组的独立转化子相比，这种SVori转化子在培养中的寿命显著延长，但它们仍然经历了衰老。我们克隆了一个转化子(SV/HF-5)的六个永生化衍生物。生长研究表明，永生化细胞系并不总是比SV/HF-5或HS74生长得更好。遗传学研究包括核型分析和整合病毒序列的Southern分析，证明了随机和非随机变化。所有永生化的衍生品都保存了表达截短T抗原的SV40序列的两个拷贝中的一个。这些克隆的SV40转化细胞系，死亡前和死亡后，应该在定义与永生化相关的分子变化方面有用。
Simian virus 40 (SV40)-mediated transformation of human diploid fibroblasts has provided an effective experimental system for studies of both “senescence” in cell culture and carcinogenesis. Previous interpretations may have been complicated, however, by the semipermissive virus-cell interaction. In earlier studies, we previously demonstrated that the human diploid fibroblast line HS74 can be efficiently transformed by DNA from replication-defective mutants of SV40 containing a deletion in the viral origin for DNA synthesis (SVori-). In the current study, we found that such SVori-transformants show a significantly increased life span in culture, as compared with either HS74 or an independent transformant containing an intact viral genome, but they nonetheless undergo senescence. We have clonally isolated six immortalized derivatives of one such transformant (SV/HF-5). Growth studies indicate that the immortalized cell lines do not invariably grow better than SV/HF-5 or HS74. Genetic studies involving karyotypic analysis and Southern analysis of integrated viral sequences demonstrated both random and nonrandom alterations. All immortalized derivatives conserved one of the two copies of SV40 sequences which expressed a truncated T antigen. These cloned SV40-transformed cell lines, pre- and postimmortalization, should be useful in defining molecular changes associated with immortalization.