脱氧胞苷激酶调控电离辐射诱导三阴性乳腺癌细胞铁死亡  

Deoxycytidine kinase participates in the regulation of ionizing radiation induced ferroptosis intriple-negative breast cancer cells

作　　者：魏刚[1] 杨永净[2] 刘博宇[3] 朱宣辑 钟睿 Wei Gang;Yang Yongjing;Liu Boyu;Zhu Xuanji;Zhong Rui(The Second Department of Breast Surgery,Jilin Cancer Hospital,Changchun 130000,China;The First Department of Radiation Oncology,Jilin Cancer Hospital,Changchun 130000,China;Department of Radiotherapy,Jilin Cancer Hospital,Changchun 130000,China;Medical Department(Medical Record),The First Hospital of Jilin University,Changchun 130021,China;Translational Cancer Research Lab,Jilin Cancer Hospital,Changchun 130000,China)

机构地区：[1]吉林省肿瘤医院乳腺外二科,长春130000 [2]吉林省肿瘤医院放疗一科,长春130000 [3]吉林省肿瘤医院放射治疗科,长春130000 [4]吉林大学白求恩第一医院医务部(病案),长春130021 [5]吉林省肿瘤医院肿瘤转化医学实验室,长春130000

出　　处：《中华放射肿瘤学杂志》2023年第12期1076-1084,共9页Chinese Journal of Radiation Oncology

基　　金：吉林省发改委项目(2021C043-1);吉林省卫健委基金(2021JC095);吉林省科技厅项目(YDZJ202301ZYTS512)。

摘　　要：目的探索脱氧胞苷激酶(dCK)在放射诱导三阴性乳腺癌(TNBC)铁死亡中的调控作用。方法用人乳腺癌细胞株MDA-MB-231构建dCK基因沉默和不同磷酸化表型的细胞模型, 并给予铁死亡诱导剂Erastin和/或铁死亡抑制剂Fer-1联合或不联合X线放射处理。通过MTT法检测细胞活性, 活性氧荧光探针(DCFH-DA)检测活性氧水平, 通过蛋白质印迹法(Western blot)检测dCK、转铁蛋白、转铁蛋白受体(TfR1)、铁转运蛋白(FPN)、铁蛋白重链1(FTH1)的蛋白表达量。采用SPSS 17.0和Origin 2021软件对数据进行分析。计量资料符合正态分布, 以x±s表示。两组之间的比较使用Studentt检验进行, 而三组及以上的比较使用单向方差分析。结果在MDA-MB-231细胞中, 放射诱导细胞死亡, 铁死亡诱导剂Erastin显著促进放射诱导的细胞死亡, 铁死亡抑制剂Fer-1能够逆转放射诱导的细胞死亡。与对照细胞相比, dCK基因沉默细胞的放射诱导的细胞死亡增加, 活性氧水平降低, Erastin联合放射诱导的细胞死亡减少, 活性氧水平减弱, Fer-1可使放射诱导的细胞死亡程度降低, 并且Fer-1无法抑制放射对活性氧的诱导作用。与对照细胞相比, 在野生型dCK(dCK-WT)或dCK过磷酸化(dCK-S74E)的dCK基因沉默细胞中, 放射诱导细胞死亡减少, 活性氧水平降低, FTH1表达量降低, 加入放射进一步降低FTH1的表达水平。此外, 在这些细胞中, Erastin促进放射诱导的细胞死亡和活性氧水平增加, Fer-1对放射诱导活性氧和细胞死亡的逆转程度明显增强。结论 dCK磷酸化促进了放射诱导TNBC细胞的铁死亡, 靶向dCK可能是一种克服TNBC治疗中辐射抗性的新治疗方式。Objective To investigate the regulatory effect of deoxycytidine kinase(dCK)on ionizing radiation(IR)induced ferroptosis in triple-negative breast cancer(TNBC).Methods TNBC cell line MDA-MB-231 was used to establish dCK knockdown and different phosphorylation phenotypes cell models,and were treated with ferroptosis activator Erastin and/or ferroptosis inhibitor ferrostatin-1(Fer-1)combined with/without X-ray irradiation.Cell viability was detected by MTT assay.The level of reactive oxygen species(ROS)was measured by 2',7'-dichlorofluorescin diacetate(DCFH-DA)staining.The expression levels of dCK,transferrin,transferrin receptor(TfR1),ferroportin(FPN)and ferritin heavy chain 1(FTH1)were detected by Western blot.Statistical analysis was performed by SPsS 17.0 and Origin 2021 software.Measurement data with normal distribution were expressed by Mean+SD.The comparison between two groups was conducted by Student t-test.The comparison among three or more groups was performed by one-way analysis of variance.Results In MDA-MB-231 cells,IR induced cell death was observed and Erastin significantly promoted radiation induced cell death,while Fer-1 was able to reverse radiation induced cell death.Compared with the control group,IR induced cell death was increased,the level of ROS was suppressed in the dCK knockdown group.Erastin combined with IR induced reduced cell death and weakened ROs level.Fer-1 reduced the degree of IR induced cell death,and it could not inhibit the induction of ROs by IR.Compared with the control cells,the rate of cell death was decreased induced by IR,the level of ROS was decreased,the expression of FTH1 was down-regulated after IR in the dCK wild-type(dCK-WT)or dCK hyperphosphorylated(dCK-S74E)MDA-MB-231 cells.In addition,Erastin promotes IR induced cell death and increased ROS levels,while Fer-1 significantly enhances the degree of reversal of IR induced ROS and cell death in these cells.Conclusions dCK phosphorylation increases ferroptosis induced by IR in TNBC cells.Targeting dCK may be a novel the

关 键 词：辐射 电离 脱氧胞苷激酶 铁死亡 三阴性乳腺癌 磷酸化 

分 类 号：R737.9[医药卫生—肿瘤]