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

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.