PARP-1 和 NF-κB 在胆汁诱导的下咽细胞 DNA 损伤和致癌表型中的作用。
The Role of PARP-1 and NF-κB in Bile-Induced DNA Damage and Oncogenic Profile in Hypopharyngeal Cells.
机构信息
The Yale Larynx Laboratory, Department of Surgery, Section of Otolaryngology, Yale School of Medicine, New Haven, Connecticut, USA.
出版信息
Laryngoscope. 2023 May;133(5):1146-1155. doi: 10.1002/lary.30284. Epub 2022 Jul 6.
OBJECTIVES/HYPOTHESIS: We recently documented that acidic bile, a gastroesophageal reflux content, can cause invasive hypopharyngeal squamous cell carcinoma, by inducing widespread DNA damage and promoting nuclear factor kappa B (NF-κB)-related oncogenic molecular events. Poly or adenosine diphosphate (ADP)-ribose polymerase-1 (PARP-1), a sensitive sensor of DNA damage, may interact with NF-κB. We hypothesized that PARP-1 is activated in hypopharyngeal cells (HCs) with marked DNA damage caused by acidic bile, hence there is an association between PARP-1 and NF-κB activation or its related oncogenic profile, in this process.
STUDY DESIGN
In vitro study.
METHODS
We targeted PARP-1 and NF-κB(p65), using pharmacologic inhibitors, 1.0 μM Rucaparib (AG014699) and 10 μM BAY 11-7082 {3-[4=methylphenyl)sulfonyl]-(2E)-propenenitrile}, respectively, or silencing their gene expression (siRNAs) and used immunofluorescence, luciferase, cell viability, direct enzyme-linked immunosorbent assays, and qPCR analysis to detect the effect of targeting PARP-1 or NF-κB in acidic bile-induced DNA damage, PARP-1, p-NF-κB, and B-cell lymphoma 2 (Bcl-2) expression, as well as NF-κB transcriptional activity, cell survival, and mRNA oncogenic phenotype in HCs.
RESULTS
We showed that (i) PARP-1 is overexpressed by acidic bile, (ii) targeting NF-κB adequately prevents the acidic bile-induced DNA double-strand breaks (DSBs) by gamma H2A histone family member X (γH2AX), oxidative DNA/RNA damage, PARP-1 overexpression, anti-apoptotic mRNA phenotype, and cell survival, whereas (iii) targeting PARP-1 preserves elevated DNA damage, NF-κB activation, and anti-apoptotic phenotype.
CONCLUSION
We document for the first time that the activation of PARP-1 is an early event during bile reflux-related head and neck carcinogenesis and that NF-κB can mediate DNA damage and PARP-1 activation. Our data encourage further investigation into how acidic bile-induced activated NF-κB mediates DNA damage in hypopharyngeal carcinogenesis.
LEVEL OF EVIDENCE
NA Laryngoscope, 133:1146-1155, 2023.
目的/假设：我们最近的研究记录表明，酸性胆汁，一种胃食管反流内容物，可通过诱导广泛的 DNA 损伤和促进核因子 kappa B（NF-κB）相关致癌分子事件，导致侵袭性咽鳞癌。多聚或腺苷二磷酸（ADP）-核糖聚合酶-1（PARP-1）是 DNA 损伤的敏感传感器，可能与 NF-κB 相互作用。我们假设在酸性胆汁引起的明显 DNA 损伤的咽细胞（HCs）中，PARP-1 被激活，因此在这个过程中，PARP-1 与 NF-κB 激活或其相关的致癌谱之间存在关联。
研究设计
体外研究。
方法
我们使用药理学抑制剂 1.0μM 鲁卡帕利（AG014699）和 10μM BAY 11-7082{3-[4=甲基苯基]磺酰基}-（2E）-丙烯腈，分别靶向 PARP-1 和 NF-κB（p65），或沉默其基因表达（siRNA），并使用免疫荧光、荧光素酶、细胞活力、直接酶联免疫吸附测定和 qPCR 分析来检测靶向 PARP-1 或 NF-κB 在酸性胆汁诱导的 DNA 损伤、PARP-1、p-NF-κB 和 B 细胞淋巴瘤 2（Bcl-2）表达以及 NF-κB 转录活性、细胞存活和 HCs 中 mRNA 致癌表型中的作用。
结果
我们表明，（i）酸性胆汁过度表达 PARP-1，（ii）靶向 NF-κB 可以充分防止酸性胆汁诱导的 DNA 双链断裂（DSBs）通过γH2A 组蛋白家族成员 X（γH2AX）、氧化 DNA/RNA 损伤、PARP-1 过表达、抗凋亡 mRNA 表型和细胞存活，而（iii）靶向 PARP-1 可维持升高的 DNA 损伤、NF-κB 激活和抗凋亡表型。
结论
我们首次证明，PARP-1 的激活是胆汁反流相关头颈部致癌发生过程中的早期事件，NF-κB 可以介导 DNA 损伤和 PARP-1 激活。我们的数据鼓励进一步研究酸性胆汁诱导的激活 NF-κB 如何介导咽癌发生中的 DNA 损伤。
证据水平
无。喉科学，133：1146-1155，2023。
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