NU7026抑制DNA-PKcs对辐射旁效应中L-02细胞增殖、凋亡、抗氧化的影响  

Influences of NU7026 on proliferation,apoptosis and anti-oxidative ability of L-02 cells in radiation-induced bystander effects by inhibiting DNA-PKcs

作　　者：万丹婷 周洁 朱子豪 邓蒸蒸 雷鹏渊 黄波 WAN Danting;ZHOU Jie;ZHU Zihao;DENG Zhengzheng;LEI Pengyuan;HUANG Bo(School of Public Health,Hengyang Medical School,University of South China,Hengyang 421001,Hunan,China)

机构地区：[1]南华大学衡阳医学院公共卫生学院,湖南衡阳421001

出　　处：《中南医学科学杂志》2023年第4期481-485,536,共6页Medical Science Journal of Central South China

基　　金：国家自然科学基金项目(81272994);湖南省教育厅科学研究重点项目(19A429);湖南省自然科学基金面上项目(2021JJ30592);湖南省卫生健康委科研计划项目(D202309037942)。

摘　　要：目的探讨NU7026抑制DNA-PKcs对辐射旁效应中L-02细胞增殖、凋亡、抗氧化的影响。方法构建辐射旁效应模型。实验分为空白组(阴性对照)、旁效应组(辐射条件培养液处理)、联合组(辐射条件培养液+NU7026)、辐照组(直接辐照)。采用MTT法、克隆形成实验、微核实验、活性氧(ROS)试剂盒、抗氧化活性试剂盒、荧光分光光度法、流式细胞术、Western blotting分别检测细胞存活率、克隆形成率、微核率、活性氧水平、超氧化物歧化酶(SOD)、谷胱甘肽(GSH)活性、丙二醛(MDA)含量、线粒体膜电位、细胞凋亡和周期阻滞及相关蛋白的表达。结果与空白组比较,其他3组的细胞存活率、克隆形成率、SOD和GSH活力、线粒体膜电位降低(P<0.05);微核率、MDA、ROS含量、凋亡率及G2/M期阻滞升高(P<0.05)。与旁效应组比较,联合组、辐照组的细胞存活率、克隆形成率、SOD和GSH活力、线粒体膜电位降低(P<0.05);微核率、MDA、ROS含量、凋亡率及G2/M期阻滞升高(P<0.05)。结论NU7026抑制DNA-PKcs能降低辐射旁效应中L-02细胞增殖、抗氧化和抗凋亡的能力。Aim To investigate the effect of NU7026 on the proliferation,apoptosis and antioxidation of L-02 cells in radiation-induced bystander effects(RIBE)by inhibiting DNA-PKcs.Methods The radiation-induced bystander effects model was established.The experiment was divided into four groups:blank group(negative control),bystander effect group(treated with irradiated conditioned medium),combination group(treated with irradiated conditioned medium and NU7026),and irradiation group(directly irradiated).MTT,clonogenic assay,micronucleus assay,reactive oxygen species(ROS)assay,antioxidant activity assay,fluorescence spectrophotometry,flow cytometry,and Western blotting were used to detect the cell survival rate,colony formation rate,micronucleus rate,ROS levels,superoxide dismutase(SOD),glutathione(GSH),malondialdehyde(MDA)content,mitochondrial membrane potential,cell apoptosis,cycle arrest,and expression of related proteins.Results Compared with the blank group,the cell survival rate,clone formation rate,SOD activity,GSH level,and mitochondrial membrane potential in the other three groups were decreased(P<0.05);The micronucleus rate,MDA,ROS content,apoptosis rate,and G2/M phase arrest were increased(P<0.05).Compared with the bystander effect group,the cell survival rate,clone formation rate,SOD activity,GSH level,and mitochondrial membrane potential in the combined group and the irradiation group were decreased(P<0.05);The micronucleus rate,MDA,ROS content,apoptosis rate,and G2/M phase arrest were increased(P<0.05).Conclusion Inhibition of DNA-PKcs by NU7026 reduced the ability of proliferation,anti-oxidative ability and anti-apoptosis of L-02 cells in radiation-induced bystander effects.

关 键 词：NU7026 DNA-PKCS 辐射旁效应 L-02细胞 细胞增殖 

分 类 号：Q691.5[生物学—生物物理学] R811.5[医药卫生—放射医学]