Zhen W, Link C J, O'Connor P M, Reed E, Parker R, Howell S B, Bohr V A
Laboratory of Molecular Pharmacology, National Cancer Institute, Bethesda, Maryland 20892.
Mol Cell Biol. 1992 Sep;12(9):3689-98. doi: 10.1128/mcb.12.9.3689-3698.1992.
We have studied several aspects of DNA damage formation and repair in human ovarian cancer cell lines which have become resistant to cisplatin through continued exposure to the anticancer drug. The resistant cell lines A2780/cp70 and 2008/c13*5.25 were compared with their respective parental cell lines, A2780 and 2008. Cells in culture were treated with cisplatin, and the two main DNA lesions formed, intrastrand adducts and interstrand cross-links, were quantitated before and after repair incubation. This quantitation was done for total genomic lesions and at the level of individual genes. In the overall genome, the initial frequency of both cisplatin lesions assayed was higher in the parental than in the derivative resistant cell lines. Nonetheless, the total genomic repair of each of these lesions was not increased in the resistant cells. These differences in initial lesion frequency between parental and resistant cell lines were not observed at the gene level. Resistant and parental cells had similar initial frequencies of intrastrand adducts and interstrand cross-links in the dihydrofolate reductase (DHFR) gene and in several other genes after cisplatin treatment of the cells. There was no increase in the repair efficiency of intrastrand adducts in the DHFR gene in resistant cell lines compared with the parental partners. However, a marked and consistent repair difference between parental and resistant cells was observed for the gene-specific repair of cisplatin interstrand cross-links. DNA interstrand cross-links were removed from three genes, the DHFR, multidrug resistance (MDR1), and delta-globin genes, much more efficiently in the resistant cell lines than in the parental cell lines. Our findings suggest that acquired cellular resistance to cisplatin may be associated with increased gene-specific DNA repair efficiency of a specific lesion, the interstrand cross-link.
我们研究了人类卵巢癌细胞系中DNA损伤形成和修复的几个方面,这些细胞系通过持续暴露于抗癌药物顺铂而对其产生了抗性。将耐药细胞系A2780/cp70和2008/c13*5.25与其各自的亲本细胞系A2780和2008进行了比较。培养的细胞用顺铂处理,在修复孵育前后对形成的两种主要DNA损伤——链内加合物和链间交联进行定量。这种定量是针对总基因组损伤以及单个基因水平进行的。在整个基因组中,所检测的两种顺铂损伤的初始频率在亲本细胞系中高于衍生的耐药细胞系。尽管如此,耐药细胞中这些损伤的总基因组修复并未增加。亲本细胞系和耐药细胞系之间在初始损伤频率上的这些差异在基因水平上未观察到。在用顺铂处理细胞后,耐药细胞系和亲本细胞系在二氢叶酸还原酶(DHFR)基因以及其他几个基因中的链内加合物和链间交联的初始频率相似。与亲本细胞系相比,耐药细胞系中DHFR基因的链内加合物修复效率没有增加。然而,在顺铂链间交联的基因特异性修复方面,观察到亲本细胞系和耐药细胞系之间存在明显且一致的修复差异。耐药细胞系比亲本细胞系更有效地从三个基因——DHFR、多药耐药(MDR1)和δ-珠蛋白基因中去除DNA链间交联。我们的研究结果表明,细胞对顺铂获得性耐药可能与特定损伤——链间交联的基因特异性DNA修复效率增加有关。