Department of Environmental Health, West China School of Public Health, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Environ Mol Mutagen. 2012 Jun;53(5):325-33. doi: 10.1002/em.21697. Epub 2012 May 10.
Deregulated expression of DNA polymerase beta (pol β) has been implicated in genomic instability that leads to tumorigenesis, yet the mechanisms underlying the pol β-mediated genetic instability remain elusive. In this study, we investigated the roles of deregulated expression of pol β in spontaneous and xenobiotic-induced genetic instability using mouse embryonic fibroblasts (MEFs) that express distinct pol β levels (wild-type, null, and overexpression) as a model system. Three genetic instability endpoints, DNA strand breaks, chromosome breakage, and gene mutation, were examined under various expression levels of pol β by comet assay, micronuclei test, and hprt mutation assay. Our results demonstrate that neither pol β deficiency nor pol β overexpression is sufficient for accumulation of spontaneous DNA damage that promotes a hyperproliferation phenotype. However, pol β null cells exhibit increased sensitivity to exogenous DNA damaging agents with increased genomic instability compared with pol β wild-type and overexpression cells. This finding suggests that a pol β deficiency may underlie genomic instability induced by exogenous DNA damaging agents. Interestingly, pol β overexpression cells exhibit less chromosomal or DNA damage, but display a higher hprt mutation frequency upon methyl methanesulfonate exposure compared with the other two cell types. Our results therefore indicate that an excessive amount of pol β may promote genomic instability, presumably through an error-prone repair response, although it enhances overall BER capacity for induced DNA damage.
DNA 聚合酶 β(pol β)的表达失调与导致肿瘤发生的基因组不稳定性有关,但 pol β 介导的遗传不稳定性的机制仍不清楚。在这项研究中,我们使用表达不同 pol β 水平(野生型、缺失型和过表达型)的小鼠胚胎成纤维细胞(MEFs)作为模型系统,研究了 pol β 表达失调在自发性和外源性遗传不稳定性中的作用。通过彗星试验、微核试验和 hprt 突变试验,在不同的 pol β 表达水平下,我们检测了三种遗传不稳定性终点,即 DNA 链断裂、染色体断裂和基因突变。我们的结果表明,pol β 缺乏或过表达本身不足以积累促进过度增殖表型的自发 DNA 损伤。然而,与 pol β 野生型和过表达细胞相比,pol β 缺失细胞对体外 DNA 损伤剂更敏感,基因组不稳定性增加。这一发现表明,pol β 缺乏可能是外源性 DNA 损伤剂诱导的基因组不稳定性的基础。有趣的是,与其他两种细胞类型相比,pol β 过表达细胞在暴露于甲基甲磺酸时表现出较少的染色体或 DNA 损伤,但 hprt 突变频率更高。因此,我们的结果表明,过量的 pol β 可能通过易错修复反应促进基因组不稳定性,尽管它增强了诱导 DNA 损伤的整体 BER 能力。