Abstract
Environmental chemical exposure often causes DNA damage, which leads to cellular dysfunction and the development of diseases. 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco-specific carcinogen that is known to cause DNA damage, while remains unknown about the underlying mechanism. In this study, simulated doses of NNK exposure in smokers, ranging from 50 to 300 μM, were used to detect the DNA damage effects of NNK in two human bronchial epithelial cells, 16HBE and BEAS-2B. The comet assay revealed increased DNA damage in response to NNK treatment, as measured by increased Olive tail moment (OTM). NNK treatment also led to elevated foci formation and protein expression of γ-H2AX, a DNA damage sensor. Dysregulation of proliferation, cell cycle arrest and apoptosis, was also observed in NNK-treated cells. Furthermore, the most effective dose of NNK (300 μM) was used in subsequent mechanistic studies. A circular RNA circNIPBL was identified to be significantly up-regulated in NNK-treated cells, circNIPBL knockdown successfully alleviated NNK-induced DNA damage and reversed the cellular dysregulation, while circNIPBL overexpression had the opposite effect. Mechanistically, we identified an interaction between circNIPBL and PARP1, a critical enzyme of the base excision repair (BER) pathway. CircNIPBL silencing successfully alleviated the NNK-induced inhibition of BER pathway proteins, including PARP1, XRCC1, PCNA and FEN1, while overexpression of circNIPBL had the opposite effect. In summary, our study shows for the first time that circNIPBL promotes NNK-induced DNA damage and cellular dysfunction through the BER pathway. In addition, our findings reveal the crucial role of epigenetic regulation in carcinogen-induced genetic lesions and further our understanding of environmental carcinogenesis.
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Funding
This study was supported by the National Natural Science Foundation of China (82130095, 91643204, 81872652 to J.Y.), Guangdong Natural Science Foundation (2018B030311019 to J.Y.), Open Research Fund of the Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan People's Hospital (202011-108 to J.Y.), and the Key Medical Disciplines and Specialties Program of Guangzhou (2021-2023).
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YL: conceptualization, methodology, data curation, validation, visualization, writing—original draft. QH: conceptualization, investigation, writing—original draft. ML: validation, methodology. XL: validation, methodology. WC: methodology. HZ: methodology. QD: methodology. CS: software, formal analysis. YL: investigation. YJ: resources, supervision, writing—review and editing, project administration, funding acquisition.
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Liu, Y., Hua, Q., Li, M. et al. Circular RNA circNIPBL promotes NNK-induced DNA damage in bronchial epithelial cells via the base excision repair pathway. Arch Toxicol 96, 2049–2065 (2022). https://doi.org/10.1007/s00204-022-03297-z
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DOI: https://doi.org/10.1007/s00204-022-03297-z