Evofosfamide Enhances Sensitivity of Breast Cancer Cells to Apoptosis and Natural-Killer-Cell-Mediated Cytotoxicity Under Hypoxic Conditions.

IF 4.5 2区 医学 Q1 ONCOLOGY
Cancers Pub Date : 2025-06-14 DOI:10.3390/cancers17121988
Shubhankar Das, Goutham Hassan Venkatesh, Walid Shaaban Moustafa Elsayed, Raefa Abou Khouzam, Ayda Shah Mahmood, Husam Hussein Nawafleh, Nagwa Ahmed Zeinelabdin, Rania Faouzi Zaarour, Salem Chouaib
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引用次数: 0

Abstract

Background/objectives: Hypoxia in the tumor microenvironment is linked to aggressiveness, epithelial-mesenchymal transition, metastasis, and therapy resistance. Targeting hypoxia to enhance antitumor immunity is crucial for overcoming therapeutic resistance. Here, we investigated the ability of Evofosfamide, a prodrug that gets activated under hypoxic conditions, to sensitize breast cancer cells to cell death. Evofosfamide is converted into bromo-isophosphoramide mustard, a potent DNA cross-linking agent that is expected to enhance the killing of cancer cells under hypoxic conditions, where these cells typically exhibit resistance. Methods: Representative breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with Evofosfamide under normoxia and hypoxia. Changes in cell viability and the mechanism of cell death were measured using neutral red dye uptake, Annexin-FITC/propidium iodide staining, and Western blot analysis of markers-PARP1 and caspase 3/7. We tested Evofosfamide's ability to counteract hypoxic suppression of type I Interferon signaling genes using quantitative PCR (qPCR), as well as its capacity to trigger natural killer (NK)-cell-mediated cytotoxicity. Results: Evofosfamide enhanced cell killing in both MCF-7 and MDA-MB-231 cells under hypoxic conditions compared to normoxic conditions. Cell killing was accompanied by increased cellular reactive oxygen species (ROS), diminished mitochondrial membrane potential, and induction of apoptosis, as demonstrated by the fragmentation or laddering of genomic DNA, the activation of caspase 3/7, and the cleavage of PARP. qPCR analysis revealed that Evofosfamide was capable of restoring type I interferon signaling in hypoxic breast cancer cells, leading to the subsequent cytolytic activity of NK cells against the tumor cells. Conclusions: Thus, conditioning the breast cancer cells with Evofosfamide resulted in enhanced cell killing under hypoxia, further underscoring its potential as a sensitizer to target hypoxia-driven tumors.

在缺氧条件下,Evofosfamide增强乳腺癌细胞对凋亡和自然杀伤细胞介导的细胞毒性的敏感性。
背景/目的:肿瘤微环境中的缺氧与侵袭性、上皮-间质转化、转移和治疗抵抗有关。靶向缺氧增强抗肿瘤免疫是克服治疗耐药的关键。在这里,我们研究了Evofosfamide(一种在缺氧条件下被激活的前药)使乳腺癌细胞对细胞死亡敏感的能力。环磷酰胺转化为溴异磷酰胺芥菜,这是一种有效的DNA交联剂,有望增强在缺氧条件下杀死癌细胞的能力,在缺氧条件下,这些细胞通常表现出耐药性。方法:对具有代表性的乳腺癌细胞系MCF-7和MDA-MB-231进行常氧和低氧处理。通过中性红色染料摄取、Annexin-FITC/碘化丙啶染色、标记物parp1和caspase 3/7的Western blot分析,检测细胞活力的变化和细胞死亡机制。我们使用定量PCR (qPCR)测试了Evofosfamide对抗I型干扰素信号基因缺氧抑制的能力,以及它触发自然杀伤(NK)细胞介导的细胞毒性的能力。结果:与常氧条件相比,在缺氧条件下,Evofosfamide增强了MCF-7和MDA-MB-231细胞的细胞杀伤。细胞杀伤伴随着细胞活性氧(ROS)的增加、线粒体膜电位的降低和凋亡的诱导,表现为基因组DNA的断裂或阶梯化、caspase 3/7的激活和PARP的裂解。qPCR分析显示,Evofosfamide能够恢复缺氧乳腺癌细胞中的I型干扰素信号,导致NK细胞随后对肿瘤细胞的细胞溶解活性。结论:因此,用依福环磷酰胺调节乳腺癌细胞可增强缺氧条件下的细胞杀伤,进一步强调其作为致敏剂靶向缺氧驱动肿瘤的潜力。
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
Cancers
Cancers Medicine-Oncology
CiteScore
8.00
自引率
9.60%
发文量
5371
审稿时长
18.07 days
期刊介绍: Cancers (ISSN 2072-6694) is an international, peer-reviewed open access journal on oncology. It publishes reviews, regular research papers and short communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced.
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