Bates Steven E, Zhou Ning Ye, Federico Laura E, Xia Liqun, O'Connor Timothy R
Department of Biology, Beckman Research Institute, City of Hope National Medical Center 1450 East Duarte Road, Duarte, CA 91010, USA.
Nucleic Acids Res. 2005 Apr 29;33(8):2475-85. doi: 10.1093/nar/gki542. Print 2005.
The progression of a normal cell to senescence in vivo and in vitro is accompanied by a reduction in the length of the telomeres, the chromosome capping segments at the end of each linkage group. However, overexpression of the reverse transcriptase subunit (HTERT) of the ribonucleoprotein telomerase restores telomere length and delays cellular senescence. Although some data exist in the literature with respect to survival, no molecular data have shown that DNA repair in telomerase-immortalized cells is normal. Several telomerase-immortalized human skin fibroblast cell lines were constructed from a primary human fibroblast cell line. The primary line and the telomerase-immortalized cell lines were treated with either ultraviolet (UV) radiation or dimethylsulfate (DMS). UV radiation principally produces cyclobutane pyrimidine dimers that are repaired by nucleotide excision repair, whereas DMS introduces mainly N-methylpurines repaired by base excision repair. Here, we show that repair of both types of damage in the telomerase-immortalized human skin fibroblast cell lines is identical to repair observed in normal skin fibroblasts. Thus, telomerase expression and consequent immortalization of skin fibroblasts do not alter nucleotide or base excision repair in human cells.
在体内和体外,正常细胞向衰老的转变伴随着端粒长度的缩短,端粒是每个连锁群末端的染色体帽状片段。然而,核糖核蛋白端粒酶的逆转录酶亚基(HTERT)的过表达可恢复端粒长度并延迟细胞衰老。尽管文献中有一些关于存活率的数据,但尚无分子数据表明端粒酶永生化细胞中的DNA修复是正常的。从原代人成纤维细胞系构建了几种端粒酶永生化的人皮肤成纤维细胞系。原代细胞系和端粒酶永生化细胞系用紫外线(UV)辐射或硫酸二甲酯(DMS)处理。紫外线辐射主要产生通过核苷酸切除修复来修复的环丁烷嘧啶二聚体,而硫酸二甲酯主要引入通过碱基切除修复来修复的N-甲基嘌呤。在此,我们表明端粒酶永生化的人皮肤成纤维细胞系中这两种类型损伤的修复与正常皮肤成纤维细胞中观察到的修复相同。因此,端粒酶表达以及由此导致的皮肤成纤维细胞永生化不会改变人细胞中的核苷酸或碱基切除修复。