Chen Oleg, Vovk Olena, Polishchuk Nikita, Mayevska Oksana, Shuvayeva Galyna, Demir Melike, Lukiyanchuk Vasyl, Kunz-Schughart Leoni A, Dubrovska Anna, Stasyk Oleh
Department of Cell Signaling, Institute of Cell Biology, National Academy of Sciences of Ukraine, Drahomanov Str. 14/16, 79005 Lviv, Ukraine.
Cancer Metabolism Group, Department of Cancer Research, Luxembourg Institute of Health, 1210 Luxembourg, Luxembourg.
Biomolecules. 2025 Jun 19;15(6):900. doi: 10.3390/biom15060900.
The development of acquired resistance to arginine deprivation therapy (ADT) is a major barrier to its efficacy. This study aimed to elucidate the possible mechanisms underlying the resistance to ADT. We applied repeated ADT and established a subline SAS-R9 of the human head and neck squamous cell carcinoma (HNSCC) cells semi-resistant to arginine (Arg) deprivation . This subline was compared to the parental SAS cell lines for its relative clonogenic proliferation, aggregation, adhesion, and migration capacities. The transcriptomic changes were assessed by RNA sequencing. Signaling pathway alterations were confirmed by RT-PCR and Western blotting. Relative cell radioresistance was analyzed by radiobiological clonogenic survival assay. DNA double-strand break (DSB) repair was assessed by γH2A.X foci analysis. SAS-R9 cells showed higher survival in response to ADT and radiotherapy, elevated clonogenic proliferation rate, cell-cell aggregation, and cell-matrix adhesion, along with increased epithelial-mesenchymal transition (EMT) markers and enhanced DNA DSB repair, potentially related to a more aggressive and therapy-resistant phenotype. While acute ADT has radiosensitizing potential, this new study suggests that long-term, repeated ADT is associated with cell selection and reprogramming, resulting in resistance to radiotherapy-induced DNA damage and higher tumor cell aggressiveness.
对精氨酸剥夺疗法（ADT）产生获得性耐药是其疗效的主要障碍。本研究旨在阐明ADT耐药的潜在机制。我们应用反复ADT并建立了对精氨酸（Arg）剥夺具有半抗性的人头颈部鳞状细胞癌（HNSCC）细胞亚系SAS-R9。将该亚系与亲代SAS细胞系比较其相对克隆增殖、聚集、黏附和迁移能力。通过RNA测序评估转录组变化。通过RT-PCR和蛋白质免疫印迹法确认信号通路改变。通过放射生物学克隆存活试验分析相对细胞放射抗性。通过γH2A.X灶点分析评估DNA双链断裂（DSB）修复。SAS-R9细胞对ADT和放疗表现出更高的存活率、克隆增殖率升高、细胞间聚集和细胞-基质黏附增加，同时上皮-间质转化（EMT）标志物增加且DNA DSB修复增强，这可能与更具侵袭性和抗治疗表型有关。虽然急性ADT具有放射增敏潜力，但这项新研究表明，长期反复ADT与细胞选择和重编程有关，导致对放疗诱导的DNA损伤产生抗性以及肿瘤细胞侵袭性增加。
