某些口腔癌患者经历放射治疗失败的事实意味着，在治疗过程中，存活癌细胞可能会出现更具放射抗性和侵袭性的表型。我们的研究旨在通过模拟临床相关放疗剂量策略的分次照射方案建立抗辐射口腔癌细胞，并研究恶性表型的全面改变。抗辐射口腔癌细胞是通过将 Cal27 和 Detroit 562 细胞暴露于 60 Gy 辐射而产生的分为 10 个剂量递增部分，并通过细胞免疫荧光进行验证。通过蛋白质印迹法评估与上皮间质转化（EMT）过程和癌症干细胞（CSC）表型相关的特异性标志物。分别使用基质胶包被的 Transwell 和伤口愈合测定来评估细胞侵袭和迁移。非靶向代谢组学用于机械地描述与 EMT 和 CSC 相关的潜在代谢模式；还通过球体形成试验和细胞免疫荧光检查了CSC表型。成功建立并验证了放射抗性口腔癌细胞系。这些细胞表现出增强的 EMT 以及细胞侵袭和迁移的增加。这些抗辐射细胞进一步表现出高代谢特征，特别是脂质代谢重编程和 ATP 结合盒 (ABC) 转运蛋白的功能富集。一致地，通过干性标志物的表达升高和球体形成能力的增加证实了放射抗性细胞中CSC表型的增强。接受分次辐射的放射抗性口腔癌细胞表现出增强的恶性表型。与增强的 EMT 和 CSC 表型相关的代谢特征为改善口腔癌放射治疗提供了潜在目标。版权所有 © 2024 Elsevier Ltd。保留所有权利。

The fact that certain oral carcinoma patients experience radiotherapy failure implies that a more radioresistant and aggressive phenotype of surviving cancer cells potentially occurs during treatment. Our study aimed to establish radioresistant oral cancer cells through a fractionated irradiation protocol that mimics clinically relevant radiotherapy dosing strategies and to investigate all-round alterations in the malignant phenotype.Radioresistant oral carcinoma cells were generated by exposing Cal27 and Detroit 562 cells to 60 Gy radiation in 10 dose-escalating fractions and verified by cell immunofluorescence. Specific markers related to the epithelial-mesenchymal transition (EMT) process and the cancer stem cell (CSC) phenotype were assessed by Western blotting. Cell invasion and migration were evaluated using Matrigel-coated transwell and wound healing assays, respectively. Nontargeted metabolomics was used to mechanistically delineate the potential metabolic patterns linked to EMT and CSCs; the CSC phenotype was also examined by sphere formation assays and cell immunofluorescence.Radioresistant oral carcinoma cell lines were successfully established and validated. These cells exhibited enhanced EMT and increase in both cell invasion and migration. These radioresistant cells further demonstrated a high metabolic profile, notably marked by lipid metabolism reprogramming and functional enrichment of ATP-binding cassette (ABC) transporters. Consistently, enhanced CSC phenotype in radioresistant cells was confirmed by elevated expression of stemness markers and increased sphere-forming capacity.Radioresistant oral carcinoma cells subjected to fractionated radiation exhibit an augmented malignant phenotype. The metabolic characteristics linked to enhanced EMT and CSC phenotypes provide potential targets for improving radiotherapy in oral carcinoma.Copyright © 2024 Elsevier Ltd. All rights reserved.