Cloning of cDNAs with possible association with senescence and immortalization of human cells

Mutation Research/DNAging Pub Date : 1994-02-01 DOI:10.1016/0921-8734(94)90005-1

Yasuhiro Satoh , Masamichi Kashimura , Shigeru Kaneko , Yuji Karasaki , Ken Higashi , Sadao Gotoh

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引用次数: 16

Abstract

Normal human diploid fibroblasts (HDF) have a finite life span in vitro and have used as a model system for the study of in vivo aging. Little in known about how changes in gene expression may affect the immortalization of human fibroblasts. We looked for cDNA clones whose mRNAs were differentially expressed between mortal senescent SV40-transformed human fibroblasts (B-32) and the immortal counterparts (B-32F) derived from B-32 cells. We identified three cDNA isolates by subtractive differential hybridization with ³²P-labeled cDNA probes from B-32 cells and B-32F cells. Nucleotide sequence analysis of these cDNA clones revealed that they wer homologous to the human vimentin, a human mitochondrial gene and a human gene of unknown nature. Slot blot and Northern blot analyses demonstrated that the former two were preferentially expressed in senescent B-32 cells and the last one was less expressed in B-32F immortal cells.

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克隆可能与人类细胞衰老和永生相关的cdna

正常人类二倍体成纤维细胞(HDF)在体外具有有限的寿命，已被用作体内衰老研究的模型系统。对于基因表达的变化如何影响人类成纤维细胞的永生化，我们知之甚少。我们寻找mrna在sv40转化的人类衰老成纤维细胞(B-32)和来自B-32细胞的不朽成纤维细胞(B-32F)之间表达差异的cDNA克隆。我们用32p标记的cDNA探针从B-32细胞和B-32F细胞中分离出3个cDNA分离株。对这些cDNA克隆进行核苷酸序列分析，发现它们分别与人类vimentin、人类线粒体基因和人类未知基因同源。Slot blot和Northern blot分析表明，前两个基因在衰老B-32细胞中优先表达，后一个基因在B-32F不朽细胞中表达较少。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Mutation Research/DNAging

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