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使用碱性彗星试验检测暴露于UVC、UVB、UVA和γ射线的人成纤维细胞中的DNA修复缺陷。

Use of the alkaline comet assay to detect DNA repair deficiencies in human fibroblasts exposed to UVC, UVB, UVA and gamma-rays.

作者信息

Alapetite C, Wachter T, Sage E, Moustacchi E

机构信息

URA 1292 du CNRS, Institut Curie-Section de Recherche, Institut Curie, Paris, France.

出版信息

Int J Radiat Biol. 1996 Mar;69(3):359-69. doi: 10.1080/095530096145922.

Abstract

The alkaline single cell gel electrophoresis (comet) assay applied to human fibroblasts allowed us to analyze the response to components of the solar spectrum (UVB and UVA) in comparison with the well-established response to UVC and gamma-rays. DNA strand breaks related to nucleotide excision repair of DNA photoproducts were produced 1 h after exposure to UVB or UVC in the normal cell line but not in the repair deficient XPD and TTD-2 cell lines. In contrast, the immediate production of DNA strand breaks observed in all cell lines after exposure to UVA or gamma-rays was followed by restitution of high molecular weight DNA upon post-exposure incubation. These results imply that (1) fibroblasts as well as lymphocytes can be analysed by the comet assay and (2) the comet assay clearly distinguishes cellular nucleotide excision repair capacity without the use of inhibitors of DNA synthesis.

摘要

将碱性单细胞凝胶电泳(彗星试验)应用于人类成纤维细胞,使我们能够分析其对太阳光谱成分(UVB和UVA)的反应,并与对UVC和γ射线已明确的反应进行比较。在正常细胞系中,暴露于UVB或UVC 1小时后会产生与DNA光产物核苷酸切除修复相关的DNA链断裂,但在修复缺陷的XPD和TTD - 2细胞系中则不会。相反,在暴露于UVA或γ射线后,所有细胞系中均观察到DNA链断裂的立即产生,随后在暴露后孵育时高分子量DNA得以恢复。这些结果表明:(1)成纤维细胞以及淋巴细胞都可以通过彗星试验进行分析;(2)彗星试验无需使用DNA合成抑制剂就能清楚地区分细胞的核苷酸切除修复能力。

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