Holcomb Nathaniel, Goswami Mamta, Han Sung Gu, Scott Tim, D'Orazio John, Orren David K, Gairola C Gary, Mellon Isabel
Department of Toxicology and Cancer Biology, The Markey Cancer Center, University of Kentucky, Lexington, KY, United States.
Toxicology Laboratory, Department of Food Science and Biotechnology of Animal Resources, College of Animal Bioscience and Technology, Konkuk University, Seoul 05029, Republic of Korea.
DNA Repair (Amst). 2017 Apr;52:70-80. doi: 10.1016/j.dnarep.2017.02.009. Epub 2017 Feb 16.
Chronic exposure to arsenic, most often through contaminated drinking water, has been linked to several types of cancer in humans, including skin and lung cancer. However, the mechanisms underlying its role in causing cancer are not well understood. There is evidence that exposure to arsenic can enhance the carcinogenicity of UV light in inducing skin cancers and may enhance the carcinogenicity of tobacco smoke in inducing lung cancers. The nucleotide excision repair (NER) pathway removes different types of DNA damage including those produced by UV light and components of tobacco smoke. The aim of the present study was to investigate the effect of sodium arsenite on the NER pathway in human lung fibroblasts (IMR-90 cells) and primary mouse keratinocytes. To measure NER, we employed a slot-blot assay to quantify the introduction and removal of UV light-induced 6-4 photoproducts (6-4 PP) and cyclobutane pyrimidine dimers (CPDs). We find a concentration-dependent inhibition of the removal of 6-4 PPs and CPDs in both cell types treated with arsenite. Treatment of both cell types with arsenite resulted in a significant reduction in the abundance of XPC, a protein that is critical for DNA damage recognition in NER. The abundance of RNA expressed from several key NER genes was also significantly reduced by treatment of IMR-90 cells with arsenite. Finally, treatment of IMR-90 cells with MG-132 abrogated the reduction in XPC protein, suggesting an involvement of the proteasome in the reduction of XPC protein produced by treatment of cells with arsenic. The inhibition of NER by arsenic may reflect one mechanism underlying the role of arsenic exposure in enhancing cigarette smoke-induced lung carcinogenesis and UV light-induced skin cancer, and it may provide some insights into the emergence of arsenic trioxide as a chemotherapeutic agent.
长期接触砷,通常是通过受污染的饮用水,已被证实与人类的多种癌症有关,包括皮肤癌和肺癌。然而,其致癌作用的潜在机制尚不清楚。有证据表明,接触砷可增强紫外线诱发皮肤癌的致癌性,并可能增强烟草烟雾诱发肺癌的致癌性。核苷酸切除修复(NER)途径可修复不同类型的DNA损伤,包括紫外线和烟草烟雾成分产生的损伤。本研究的目的是调查亚砷酸钠对人肺成纤维细胞(IMR-90细胞)和原代小鼠角质形成细胞中NER途径的影响。为了测量NER,我们采用狭缝印迹法来量化紫外线诱导的6-4光产物(6-4PP)和环丁烷嘧啶二聚体(CPD)的引入和去除。我们发现,在亚砷酸盐处理的两种细胞类型中,6-4PP和CPD的去除均呈浓度依赖性抑制。亚砷酸盐处理这两种细胞类型均导致XPC丰度显著降低,XPC是一种对NER中DNA损伤识别至关重要的蛋白质。用亚砷酸盐处理IMR-90细胞也显著降低了几个关键NER基因表达的RNA丰度。最后,用MG-132处理IMR-90细胞消除了XPC蛋白的减少,这表明蛋白酶体参与了砷处理细胞产生的XPC蛋白的减少。砷对NER的抑制作用可能反映了砷暴露在增强香烟烟雾诱导的肺癌发生和紫外线诱导的皮肤癌中的作用机制之一,并且可能为三氧化二砷作为一种化疗药物的出现提供一些见解。