Combined effects of carbon ion radiation and PARP inhibitor on non-small cell lung carcinoma cells: Insights into DNA repair pathways and cell death mechanisms

IF 3 3区生物学 Q2 GENETICS & HEREDITY

DNA Repair Pub Date : 2024-10-19 DOI:10.1016/j.dnarep.2024.103778

Payel Dey, Rima Das, Sandipan Chatterjee, Roni Paul, Utpal Ghosh

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引用次数: 0

Abstract

The utilization of high linear energy transfer (LET) carbon ion (¹²C-ion) in radiotherapy has witnessed a notable rise in managing highly metastatic, recurrent, and chemo/radio-resistant human cancers. Non-small cell lung cancer (NSCLC) presents a formidable challenge due to its chemo-resistance and aggressive nature, resulting in poor prognosis and survival rates. In a previous study, we demonstrated that the combination of ¹²C-ion with the poly (ADP-ribose) polymerase (PARP) inhibitor (PARPi) olaparib significantly mitigated metastasis in A549 cells. Here, we delve into the underlying rationale behind the combined action of olaparib with ¹²C-ion, focusing on DNA repair pathways and cell death mechanisms in asynchronous NSCLC A549 cells following single and combined treatments. Evaluation included analysis of colony-forming ability, DNA damage assessed by γH2AX foci, expression profiling of key proteins involved in Homologous Recombination (HR) and Non-Homologous End Joining (NHEJ) repair pathways, caspase-3 activation, apoptotic body formation, and autophagic cell death. Our findings reveal that both PARPi olaparib and rucaparib sensitize A549 cells to ¹²C-ion exposure, with olaparib exhibiting superior sensitization. Moreover, ¹²C-ion exposure alone significantly downregulates both HR and NHEJ repair pathways by reducing the expression of MRE11--RAD51 and Ku70-Ku80 protein complexes at 24 h post-treatment. Notably, the combination of olaparib pre-treatment with ¹²C-ion markedly inhibits both HR and NHEJ pathways, culminating in DNA damage-induced apoptotic and autophagic cell death. Thus we are the first to demonstrate that olaparib sensitizes NSCLC cells to carbon ion by interfering with HR and NHEJ pathway. These insights underscore the promising therapeutic potential of combining PARP inhibition with carbon ion exposure for effective NSCLC management.

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碳离子辐射和 PARP 抑制剂对非小细胞肺癌细胞的联合影响：对 DNA 修复途径和细胞死亡机制的启示

在放疗中利用高线性能量转移（LET）碳离子（12C-离子）治疗高度转移、复发和化疗/放疗耐药的人类癌症的案例显著增加。非小细胞肺癌（NSCLC）因其化疗耐药性和侵袭性，导致预后和存活率较低，是一项艰巨的挑战。在之前的一项研究中，我们证实了 12C 离子与多（ADP-核糖）聚合酶（PARP）抑制剂（PARPi）奥拉帕利（olaparib）的联合用药能显著减轻 A549 细胞的转移。在此，我们深入探讨了奥拉帕利和 12C 离子联合作用的基本原理，重点研究了单药和联合用药后异步 NSCLC A549 细胞的 DNA 修复途径和细胞死亡机制。评估包括分析集落形成能力、通过γH2AX病灶评估的DNA损伤、参与同源重组（HR）和非同源末端连接（NHEJ）修复途径的关键蛋白的表达谱、Caspase-3激活、凋亡体形成和自噬细胞死亡。我们的研究结果表明，PARPi olaparib 和 rucaparib 都能使 A549 细胞对 12C 离子暴露敏感，其中 olaparib 的敏感性更强。此外，在处理后24小时，单独暴露于12C-离子可通过减少MRE11--RAD51和Ku70-Ku80蛋白复合物的表达，显著下调HR和NHEJ修复途径。值得注意的是，奥拉帕尼预处理与 12C 离子结合使用会明显抑制 HR 和 NHEJ 途径，最终导致 DNA 损伤诱导的细胞凋亡和自噬死亡。因此，我们首次证明奥拉帕利通过干扰HR和NHEJ途径使NSCLC细胞对碳离子敏感。这些见解强调了将 PARP 抑制与碳离子暴露相结合以有效治疗 NSCLC 的巨大治疗潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

DNA Repair 生物-毒理学

CiteScore

7.60

自引率

5.30%

发文量

审稿时长

59 days

期刊介绍： DNA Repair provides a forum for the comprehensive coverage of DNA repair and cellular responses to DNA damage. The journal publishes original observations on genetic, cellular, biochemical, structural and molecular aspects of DNA repair, mutagenesis, cell cycle regulation, apoptosis and other biological responses in cells exposed to genomic insult, as well as their relationship to human disease. DNA Repair publishes full-length research articles, brief reports on research, and reviews. The journal welcomes articles describing databases, methods and new technologies supporting research on DNA repair and responses to DNA damage. Letters to the Editor, hot topics and classics in DNA repair, historical reflections, book reviews and meeting reports also will be considered for publication.