癌症是一个全球性的健康问题，占全球死亡人数的近六分之一。传统疗法与新疗法相结合提高了这种毁灭性疾病的生存率。然而，治疗耐药性的持续挑战和可用于特定癌症类型的有限治疗手段仍然迫切需要研究新的治疗策略。氯喹与不同药物（Panobinostat、KU-57788 和 NU-7026）的组合在 8 年进行了测试。人源癌细胞系（结直肠癌：HCT116 和 HT29；乳腺癌：MDA-MB-231 和 HCC1937；胶质母细胞瘤：A-172 和 LN-18；头颈部：CAL-33 和 32816）。通过 MTT 测定测试药物对增殖的影响，并通过 Anexin V-PI 细胞凋亡测定评估细胞死亡。通过磷酸化-H2AX 荧光染色分析 DNA 双链断裂的存在。为了测量同源重组效率，使用了 HR-GFP 报告基因，它允许基于流式细胞术检测 I-SceI 核酸内切酶诱导的 DSB 刺激的 HR。氯喹与所用任何药物的组合对细胞凋亡诱导显示出有效的协同作用，在胶质母细胞瘤和头颈癌细胞系中观察到特别显着的功效。我们发现氯喹产生依赖于活性氧形成的DNA双链断裂，而帕比司他通过同源重组抑制DNA双链断裂修复。 N-乙酰半胱氨酸（一种活性氧清除剂）可显着减少氯喹/帕比司他组合引起的细胞死亡，这强调了 DSB 生成在 CQ/LBH 诱导的致死性中的关键作用。基于这些数据，我们还探索了 CQ 与另一种主要 DSB 修复途径非同源末端连接 (NHEJ) 的两种抑制剂 KU-57788 和 NU-7026 的组合，并再次观察到对细胞凋亡诱导的协同作用。为 CQ 与 DSB 抑制剂联合治疗不同实体瘤的临床研究提供依据。版权所有 © 2024 Iglesias-Corral, García-Valles, Arroyo-Garrapucho, Bueno-Martínez, Ruiz-Robles, Ovejero-Sánchez, González -萨米恩托和埃雷罗。

Cancer is a global health problem accounting for nearly one in six deaths worldwide. Conventional treatments together with new therapies have increased survival to this devastating disease. However, the persistent challenges of treatment resistance and the limited therapeutic arsenal available for specific cancer types still make research in new therapeutic strategies an urgent need.Chloroquine was tested in combination with different drugs (Panobinostat, KU-57788 and NU-7026) in 8 human-derived cancer cells lines (colorectal: HCT116 and HT29; breast: MDA-MB-231 and HCC1937; glioblastoma: A-172 and LN-18; head and neck: CAL-33 and 32816). Drug´s effect on proliferation was tested by MTT assays and cell death was assessed by Anexin V-PI apoptosis assays. The presence of DNA double-strand breaks was analyzed by phospho-H2AX fluorescent staining. To measure homologous recombination efficiency the HR-GFP reporter was used, which allows flow cytometry-based detection of HR stimulated by I-SceI endonuclease-induced DSBs.The combination of chloroquine with any of the drugs employed displayed potent synergistic effects on apoptosis induction, with particularly pronounced efficacy observed in glioblastoma and head and neck cancer cell lines. We found that chloroquine produced DNA double strand breaks that depended on reactive oxygen species formation, whereas Panobinostat inhibited DNA double-strand breaks repair by homologous recombination. Cell death caused by chloroquine/Panobinostat combination were significantly reduced by N-Acetylcysteine, a reactive oxygen species scavenger, underscoring the pivotal role of DSB generation in CQ/LBH-induced lethality. Based on these data, we also explored the combination of CQ with KU-57788 and NU-7026, two inhibitors of the other main DSB repair pathway, nonhomologous end joining (NHEJ), and again synergistic effects on apoptosis induction were observed.Our data provide a rationale for the clinical investigation of CQ in combination with DSB inhibitors for the treatment of different solid tumors.Copyright © 2024 Iglesias-Corral, García-Valles, Arroyo-Garrapucho, Bueno-Martínez, Ruiz-Robles, Ovejero-Sánchez, González-Sarmiento and Herrero.