BACKGROUND AND PURPOSE Radiotherapy plays a crucial role in the treatment of cervical cancer, but existing radiosensitizers have limited efficacy in clinical applications. The aims of this study were to establish and verify an efficient method for identifying new radiosensitizers, to use this to identify candidate radiosensitizers for cervical cancer, and to investigate the specific mechanisms of these when used in combination with radiotherapy. MATERIALS AND METHODS An automated platform for identifying radiosensitizers for cervical cancer was created based on high-throughput screening technology. The radiosensitizing effects of candidate compounds from the LOPAC1280 chemical library were evaluated in radiosensitive and radioresistant cervical cancer cells using a clonogenic survival assay, with cell cycle analyses, and western blot analyses performed for both cell lines. RESULTS The automated high-throughput screening approach identified four hit compounds. One of the most potent candidates was dihydroouabain (DHO), an inhibitor of Na+/K+-ATPase that has not previously been classified as a radiosensitizer. DHO significantly enhanced radiosensitivity in cervical cancer cells. It also abrogated radiation-induced S phase arrest in cervical cancer cells. Combination treatment significantly caused the inhibition of Chk1 and increased DNA double-strand breaks (DSB). CONCLUSIONS DHO is a novel radiosensitizer for the treatment of cervical cancer. The automated high-throughput screening platform developed in this study proved to be powerful and effective, with the potential to be widely used in the future identification of radiosensitizers.

中文翻译：

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双氢哇巴因，一种通过自动化高通量筛选确定的宫颈癌新型放射增敏剂

背景和目的放射治疗在宫颈癌的治疗中起着至关重要的作用，但现有的放射增敏剂在临床应用中的疗效有限。本研究的目的是建立和验证一种识别新放射增敏剂的有效方法，用它来识别宫颈癌的候选放射增敏剂，并研究这些与放疗联合使用时的具体机制。材料和方法 基于高通量筛选技术创建了一个用于识别宫颈癌放射增敏剂的自动化平台。LOPAC1280 化学文库中候选化合物的放射增敏作用在放射敏感性和抗放射性宫颈癌细胞中使用克隆存活试验和细胞周期分析进行评估，和蛋白质印迹分析对两种细胞系进行。结果 自动化高通量筛选方法鉴定了四种命中化合物。最有效的候选药物之一是二氢哇巴因 (DHO)，它是一种 Na+/K+-ATP 酶抑制剂，以前未被归类为放射增敏剂。DHO显着增强宫颈癌细胞的放射敏感性。它还消除了宫颈癌细胞中辐射诱导的 S 期停滞。联合治疗显着导致 Chk1 的抑制和 DNA 双链断裂 (DSB) 的增加。结论 DHO 是一种用于治疗宫颈癌的新型放射增敏剂。本研究开发的自动化高通量筛选平台被证明是强大而有效的，有可能在未来的放射增敏剂鉴定中得到广泛应用。结果 自动化高通量筛选方法鉴定了四种命中化合物。最有效的候选药物之一是二氢哇巴因 (DHO)，它是一种 Na+/K+-ATP 酶抑制剂，以前未被归类为放射增敏剂。DHO显着增强宫颈癌细胞的放射敏感性。它还消除了宫颈癌细胞中辐射诱导的 S 期停滞。联合治疗显着导致 Chk1 的抑制和 DNA 双链断裂 (DSB) 的增加。结论 DHO 是一种用于治疗宫颈癌的新型放射增敏剂。本研究开发的自动化高通量筛选平台被证明是强大而有效的，有可能在未来的放射增敏剂鉴定中得到广泛应用。结果 自动化高通量筛选方法鉴定了四种命中化合物。最有效的候选药物之一是二氢哇巴因 (DHO)，它是一种 Na+/K+-ATP 酶抑制剂，以前未被归类为放射增敏剂。DHO显着增强宫颈癌细胞的放射敏感性。它还消除了宫颈癌细胞中辐射诱导的 S 期停滞。联合治疗显着导致 Chk1 的抑制和 DNA 双链断裂 (DSB) 的增加。结论 DHO 是一种用于治疗宫颈癌的新型放射增敏剂。本研究开发的自动化高通量筛选平台被证明是强大而有效的，有可能在未来的放射增敏剂鉴定中得到广泛应用。