Dong H Y, Buard A, Lévy F, Renier A, Laval F, Jaurand M C
Laboratory of Environmental Cellular and Molecular Pathology, Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Henri Mondor, Institut Mondor de Médecine Moléculaire, Faculté de Médecine, Créteil, France.
Mutat Res. 1995 Oct;331(2):197-204. doi: 10.1016/0027-5107(95)00069-u.
To investigate the origin of DNA repair in rat pleural mesothelial cells (RPMC) exposed to asbestos fibers, poly(ADP-ribose) polymerase (PARP) activity was measured in the asbestos-treated cells. As bleomycin has been shown to activate poly(ADP-ribose) synthesis in several cell systems, the response to bleomycin with regard to PARP assay was first investigated. Bleomycin produced a dose-dependent increase of poly(ADP-ribose) synthesis in RPMC. Likewise both chrysotile and crocidolite fibers produced a concentration-dependent PARP activation indicating that the formation of DNA strand breaks is one type of damage produced by asbestos in RPMC. Enhancement of DNA repair, assessed by the measurement of [3H] methylthymidine incorporation in growth arrested cells, was not detectable in the presence of 3-methoxybenzamide (3-MBA), a PARP inhibitor, confirming a relation between PARP activation and DNA repair. The participation of DNA breakage in asbestos toxicity on RPMC was determined by the colorimetric 3-4(5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. There was no relationship between DNA breakage and cytotoxicity since the use of PARP inhibitors did not change cell viability. These results indicate that asbestos produce DNA damage that is repaired in RPMC.
为了研究暴露于石棉纤维的大鼠胸膜间皮细胞(RPMC)中DNA修复的起源,我们检测了经石棉处理的细胞中的聚(ADP-核糖)聚合酶(PARP)活性。由于博来霉素已被证明能在多种细胞系统中激活聚(ADP-核糖)合成,因此首先研究了RPMC对博来霉素在PARP检测方面的反应。博来霉素使RPMC中的聚(ADP-核糖)合成呈剂量依赖性增加。同样,温石棉和青石棉纤维也产生了浓度依赖性的PARP激活,表明DNA链断裂的形成是石棉在RPMC中产生的一种损伤类型。通过测量生长停滞细胞中[3H]甲基胸苷掺入来评估的DNA修复增强,在PARP抑制剂3-甲氧基苯甲酰胺(3-MBA)存在下无法检测到,这证实了PARP激活与DNA修复之间的关系。通过比色法3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)检测确定了DNA断裂在石棉对RPMC毒性中的作用。由于使用PARP抑制剂并没有改变细胞活力,因此DNA断裂与细胞毒性之间没有关系。这些结果表明石棉会产生DNA损伤,而RPMC会对其进行修复。