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PARP-1 缺陷对苯并(a)芘暴露下人支气管上皮细胞 DNA 损伤与修复的影响。

Effect of PARP-1 deficiency on DNA damage and repair in human bronchial epithelial cells exposed to Benzo(a)pyrene.

机构信息

Department of Preventive Medicine, School of Public Health, Sun Yat-sen University, 74 Zhongshan 2nd Road, Guangzhou 510080, Guangdong, People's Republic of China.

出版信息

Mol Biol Rep. 2009 Nov;36(8):2413-22. doi: 10.1007/s11033-009-9472-z. Epub 2009 Feb 27.

Abstract

Benzo[a]pyrene is a ubiquitously distributed environmental pollutant known to cause DNA damage, whereas PARP-1 is a nuclear enzyme that is activated by damaged DNA and plays an important role in base excision repair and genomic stability. Here, 16HBE and its PAPR1-deficient cells were exposed to BaP, and the DNA damage level and repair ability of both cell lines were measured by alkaline comet assay. The results showed that cell viability of both cell lines decreased in a dose-dependent manner when exposed to BaP, but there was no significant difference between two cell lines. Comet assay showed that BaP caused DNA damage in both cell lines at an obvious dose- and time-dependent manner. Compare with 16HBE, the PARP1-deficient cells were more sensitive to the damage caused by BaP. The results of DNA repair experiment showed that both cell lines can recover from the damage in a time-dependent pattern. The relative repair percentage of PARP1-deficient cells were generally lower than that of 16HBE at all exposed concentrations at the early stage of repair, but tended to be closer between two cell lines at the later period. According to results, we came to the conclusion that PARP1-deficient cells were more sensitive to BaP in contrast to normal 16HBE; DNA repair capacity in PARP1-deficient cells decreased significantly at the early stage of repair, but increased to the equivalent level of normal 16HBE in the later period. PARP-1 plays an important role in early repair of DNA damage caused by BaP in 16HBE notwithstanding the main repair work is taken by NER pathway.

摘要

苯并[a]芘是一种广泛分布的环境污染物,已知会导致 DNA 损伤,而 PARP-1 是一种核酶,可被受损 DNA 激活,在碱基切除修复和基因组稳定性中发挥重要作用。在这里,16HBE 及其缺乏 PARP1 的细胞被暴露于 BaP 下,并用碱性彗星试验测量了这两种细胞系的 DNA 损伤水平和修复能力。结果表明,两种细胞系的细胞活力均随 BaP 暴露剂量呈剂量依赖性下降,但两种细胞系之间没有显著差异。彗星试验表明,BaP 以明显的剂量和时间依赖性方式在两种细胞系中引起 DNA 损伤。与 16HBE 相比,PARP1 缺陷细胞对 BaP 引起的损伤更为敏感。DNA 修复实验的结果表明,两种细胞系均能以时间依赖性方式从损伤中恢复。在所有暴露浓度的早期修复阶段,PARP1 缺陷细胞的相对修复百分比通常低于 16HBE,但在后期两个细胞系之间趋于接近。根据结果,我们得出结论,与正常的 16HBE 相比,PARP1 缺陷细胞对 BaP 更敏感;PARP1 缺陷细胞的 DNA 修复能力在修复的早期阶段显著下降,但在后期增加到与正常 16HBE 相当的水平。PARP-1 在 16HBE 中对 BaP 引起的 DNA 损伤的早期修复中发挥重要作用,尽管 NER 途径承担主要的修复工作。

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