The spectrum of mitochondrial DNA deletions is a ubiquitous marker of ultraviolet radiation exposure in human skin
The spectrum of mitochondrial DNA deletions is a ubiquitous marker of ultraviolet radiation exposure in human skin
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DOI：
10.1046/j.1523-1747.2000.00092.x
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发表时间：
2000-10-01
影响因子：
6.5
通讯作者：
Birch-Machin, MA
中科院分区：
文献类型：
--
作者：
Ray, AJ;Turner, R;Birch-Machin, MA
我们和同事们认为，线粒体DNA的缺失可能是有用的皮肤紫外线辐射暴露的生物标志物。在这项研究中，使用西南部的方法，涉及单克隆抗体对胸腺嘧啶二聚体，我们提供了直接的证据，紫外线诱导的线粒体DNA损伤纯化的任何核DNA污染的存在。以前的研究是有限的，因为他们集中在一个单一的线粒体DNA缺失的频率。因此，我们解决了皮肤中线粒体DNA缺失谱的问题，以及这是否可以用作整体DNA损伤的指标。我们使用了长聚合酶链反应技术，以确定几乎整个线粒体基因组的线粒体DNA缺失谱在71个分裂的皮肤样本中与太阳照射有关。随着表皮中紫外线暴露量的增加，缺失数量显著增加（Kruskal-Wallis检验，p = 0.0015），但真皮中的缺失数量没有显著增加（p = 0.6376）。在表皮中的发现并没有受到任何年龄依赖性增加的线粒体DNA缺失的混淆，也通过长聚合酶链反应技术检测到。在我们的研究中发现的大范围的缺失突出了与紫外线暴露和时间老化相关的线粒体DNA的普遍存在的性质和高突变负荷。与使用竞争性聚合酶链反应检测单个缺失相比，我们发现：长聚合酶链反应是一种敏感的技术，因此可能提供一个更全面的，虽然不是定量的，皮肤中的整体线粒体DNA损伤的指数。
We and colleagues have suggested that deletions of mitochondrial DNA may be useful as a biomarker of ultraviolet radiation exposure in skin. In this study using a southwestern approach involving monoclonal antibodies against thymine dimers we provide direct evidence for the presence of ultraviolet-induced damage in mitochondrial DNA purified from any nuclear DNA contamination. Previous studies have been limited, as they have focused on the frequency of a single mitochondrial DNA deletion. Therefore we have addressed the question of the spectrum of mitochondrial DNA deletions in skin and whether this can be used as an index of overall DNA damage, We have used a long polymerase chain reaction technique to determine the mitochondrial DNA deletion spectrum of almost the entire mitochondrial genome in 71 split skin samples in relation to sun exposure. There was a significant increase in the number of deletions with increasing ultraviolet exposure in the epidermis (Kruskal-Wallis test, p = 0.0015) but not the dermis (p = 0.6376). The findings in the epidermis are not confounded by any age-dependent increases in mitochondrial DNA deletions also detected by the long polymerase chain reaction technique. The large spectrum of deletions identified in our study highlights the ubiquitous nature and the high mutational load of mitochondrial DNA associated with ultraviolet exposure and chronologic aging. Compared with the detection of single deletions using competitive polymerase chain reaction, we show that: long polymerase chain reaction is a sensitive technique and may therefore provide a more comprehensive, although not quantitative, index of overall mitochondrial DNA damage in skin.