Differential repair of UVB-induced cyclobutane pyrimidine dimers in cultured human skin cells and whole human skin
Differential repair of UVB-induced cyclobutane pyrimidine dimers in cultured human skin cells and whole human skin
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DOI：
10.1016/j.dnarep.2008.01.005
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发表时间：
2008-05-03
期刊：
影响因子：
3.8
通讯作者：
Douki, Thierry
中科院分区：
文献类型：
--
作者：
Mouret, Stephane;Charveron, Marie;Douki, Thierry
环丁烷嘧啶二聚体（CPD）和嘧啶（6-4）嘧啶酮光产物（6-4PPs）是UVB辐射诱导的两类主要的DNA突变损伤。到目前为止，已经有许多研究致力于它们在人体细胞和皮肤中的形成和修复。然而，所使用的生物化学方法通常缺乏特异性，使得能够对TT、TC、CT和CC二核苷酸产生的四种CPD和6- 4PP中的每一种进行单独研究。在目前的工作中，我们应用高效液相色谱-质谱分析，以研究形成和修复的CPD和6-4PPs的光产物在原代培养的人角质形成细胞和成纤维细胞，以及在整个人类皮肤。我们首先观察到成纤维细胞中二聚体病变的产量略高于角质形成细胞。相反，最后一种细胞类型的整体修复率较高。此外，在皮肤活检中发现DNA光产物的去除比两种培养的皮肤细胞慢。与以前的工作一致，发现在三种类型的样品中，6-4PPs的修复比CPD更有效，TT和TC衍生物的去除之间没有观察到差异。相反，观察到两种修饰的嘧啶的性质对CPD的修复率的显著影响。去除率由高到低的顺序为：C T > C C > T C > T T T.这些数据，连同已知的内在突变性质的病变，将支持报告的紫外线突变光谱。一个值得注意的例外涉及CC二核苷酸，其是具有UV特异性CC至TT串联突变的突变热点，尽管相关的联嘧啶光产物以低产量产生并有效修复。(c)2008 Elsevier B. V.保留所有权利。
Cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6-4) pyrimidone photoproducts (6-4PPs) are the two main classes of mutagenic DNA damages induced by UVB radiation. Numerous studies have been devoted so far to their formation and repair in human cells and skin. However, the biochemical methods used often lack the specificity that would allow the individual study of each of the four CPDs and 6-4PPs produced at TT, TC, CT and CC dinucleotides. In the present work, we applied an HPLC-mass spectrometry assay to study the formation and repair of CPDs and 6-4PPs photoproducts in primary cultures of human keratinocytes and fibroblasts as well as in whole human skin. We first observed that the yield of dimeric lesions was slightly higher in fibroblasts than in keratinocytes. In contrast, the rate of global repair was higher in the last cell type. Moreover, removal of DNA photoproducts in skin biopsies was found to be slower than in both cultured skin cells. In agreement with previous works, the repair of 6-4PPs was found to be more efficient than that of CPDs in the three types of samples, with no observed difference between the removal of the TT and TC derivatives. In contrast, a significant influence of the nature of the two modified pyrimidines was observed on the repair rate of CPDs. The decreasing order of removal efficiency was the following: C T > C C > T C > T T. These data, together with the known intrinsic mutational properties of the lesions, would support the reported UV mutation spectra. A noticeable exception concerns CC dinucleotides that are mutational hotspots with an UV-specific CC to TT tandem mutation, although related bipyrimidine photoproducts are produced in low yields and efficiently repaired. (c) 2008 Elsevier B.V. All rights reserved.