收稿日期: 2023-12-25
网络出版日期: 2024-05-15
基金资助
国家自然科学基金项目(82102974);广州市科技计划项目(202201011048)
Copper regulates the radiosensitivity of nasopharyngeal carcinoma by activating MAPK-ERK signaling pathway
Received date: 2023-12-25
Online published: 2024-05-15
目的 探究铜离子（Cu2+）对鼻咽癌（nasopharyngeal carcinoma，NPC）细胞放射敏感性的影响，寻找提升NPC放疗效果的潜在靶标。 方法 细胞Cu2+检测对比正常鼻咽上皮细胞系NP69及多种NPC细胞系内Cu2+的含量；分别加入0、5、10、20、50、100、200 μmol/L Cu2+溶液及0、0.05、0.1、0.2、0.5、1.0、2.0 mmol/L TEPA溶液，利用CCK?8实验测定NPC细胞内Cu2+含量对辐照前后细胞存活率的影响，并确定后续实验中Cu2+组及TEPA组所使用的药物浓度；CCK?8实验及克隆形成实验检测药物处理后各组NPC细胞放射敏感性的变化；Western blot检测各组NPC细胞辐照处理前后的DNA损伤情况及MAPK-ERK通路相关蛋白表达。 结果 NPC细胞中Cu2+含量显著高于正常鼻咽上皮细胞（P < 0.05）。添加Cu2+溶液浓度低于50 μmol/L时促进CNE1细胞的放疗抵抗性，添加0.1 ~ 0.2 mmol/L TEPA能提高SUNE1细胞放射敏感性（P < 0.05）。与对照组比较，Cu2+组NPC细胞放疗抵抗性增强，TEPA组NPC细胞放疗敏感性增强（P < 0.05）。较之对照组而言，Cu2+组细胞MAPK-ERK通路活化程度上调（P < 0.05）。使用MAPK-ERK通路抑制剂SCH772984能够有效逆转Cu2+介导的NPC放疗抵抗（P < 0.05）。 结论 NPC细胞中Cu2+含量升高，其通过激活MAPK-ERK通路增强NPC细胞的放疗抵抗性。
关键词： 鼻咽癌; 铜离子; MAPK-ERK通路; 放射治疗
黄秀婷 , 林颉 , 叶晓心 , 蔡佳佐 , 袁亚维 . 铜离子激活MAPK-ERK通路调控鼻咽癌放射敏感性[J]. 实用医学杂志, 2024 , 40(9) : 1191 -1196 . DOI: 10.3969/j.issn.1006-5725.2024.09.003
Objective To Investigate the effect of copper ions on the radiosensitivity of nasopharyngeal carcinoma （NPC） and identify potential targets to improve the efficacy of NPC radiation therapy. Methods The content of copper ions in normal nasopharyngeal epithelial cell line NP69 and a variety of NPC cell lines was detected by a copper microplate assay. After adding 0， 5， 10， 20， 50， 100， 200 μmol/L copper ion solution and 0， 0.05， 0.1， 0.2， 0.5， 1.0， 2.0 mmol/L TEPA solution， CCK?8 assay was used to determine the effect of intracellular copper ion content on the survival rate of NPC cells before and after irradiation. And determine the drug concentration to be used in subsequent experiments. CCK?8 assay was used to determine the effect of intracellular copper ion content on the survival rate of NPC cells before and after irradiation， and to determine the drug concentration used in the copper ion group and TEPA group in the subsequent experiments. CCK?8 assay and clone formation assay were used to detect the changes in radiosensitivity of NPC cells after drug treatment. The DNA damage and MAPK-ERK signaling pathway-related protein expression of NPC cells in each group before and after irradiation were detected by Western blot. Results The content of copper ions in NPC cells was significantly higher than that in normal nasopharyngeal epithelial cells （P < 0.05）. When the concentration of copper ion solution was lower than 50 μmol/L， the radiation resistance of CNE1 cells was improved， and the radiation sensitivity of SUNE1 cells was increased by 0.1 ~ 0.2 mmol/L TEPA （P < 0.05）. Compared with the control group， the NPC cell radiotherapy resistance of copper ion group was enhanced， and the NPC cell radiotherapy sensitivity of copper ion group was enhanced （P < 0.05）. Compared with the control group， the activation degree of MAPK-ERK pathway in copper ion group was up-regulated （P < 0.05）. MAPK-ERK pathway inhibitor SCH772984 effectively reversed copper-mediated NPC radiotherapy resistance （P < 0.05）. Conclusion The increased copper ion content in NPC cells promoted the radiotherapy resistance of NPC cells by activating the MAPK-ERK signaling pathway.
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