KU60019抑制ATM对HepG2细胞辐射旁效应的调控  

The regulation of KU60019 inhibiting ATM on radiation-induced bystander effects in HepG2 cells

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

机构地区：[1]南华大学衡阳医学院公共卫生学院,湖南省衡阳市421001

出　　处：《中南医学科学杂志》2022年第3期327-330,共4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)。

摘　　要：目的探讨KU60019抑制ATM对HepG2细胞辐射旁效应的调控作用。方法使用KU60019抑制ATM,通过转移辐射条件刺激液构建HepG2细胞辐射旁效应模型。实验分为空白组(NC组)、辐射旁效应组(ICM组)、辐射旁效应+KU60019组(联合组)、单纯辐照组(IR组)。MTT法检测细胞存活率;生长曲线实验检测细胞增殖;微板法检测细胞内超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量;活性氧试剂盒检测细胞内活性氧(ROS)水平;荧光分光光度法检测细胞线粒体膜电位(MMP);流式细胞术检测细胞凋亡率。结果随着KU60019浓度增加,HepG2细胞数量逐渐减少,后续选择7μmol/L KU60019进行实验。与NC组比较,ICM组、联合组与IR组的细胞存活率、增殖能力、SOD活力、线粒体的膜电位逐渐降低(P<0.05),细胞凋亡率、ROS与MDA含量逐渐升高(P<0.05)。结论KU60019抑制ATM能降低HepG2细胞抗氧化能力,提高辐射敏感性。Aim To investigate the regulation of KU60019 inhibiting ATM on radiation-induced bystander effects in HepG2 cells.Methods ATM protein kinase was inhibited by KU60019,and the radiation-induced bystander effect model was constructed by transferring radiative conditioned stimulation fluid to HepG2 cells.The experiment was divided into NC group(NC group),radiation-induced bystander effect group(ICM group),radiation-induced bystander effect+KU60019 group(combined group)and simple irradiation group(IR group).Cell viability was detected by MTT assay.Cell proliferation was detected by growth curve experiment.The viability of superoxide dismutase(SOD)and malondialde-hyde content(MDA)in cells were detected by microplate method.The levels of reactive oxygen species(ROS)was de-tected by ROS kit.Mitochondrial membrane potential(MMP)was detected by fluorescence spectrophotometry.Apoptosis was detected by flow cytometry.Results As the concentration of KU60019 increased,the number ofHep G2 cells gradually decreased,and 7μmol/L KU60019 concentration was selected for subsequent experiments.Com-pared with NC group,the cell survival rate,proliferative capacity,SOD activity and mitochondrial membrane potential ofICM group,combined group and IR group decreased gradually(P<0.05).The cell apoptosis rate,ROS and MDA con-tent increased gradually(P<0.05).Conclusion Inhibition of ATM by KU60019 decreased the antioxidant capacityof HepG2 cells and increased the radiation sensitivity.

关 键 词：共济失调毛细血管扩张症突变 ATM抑制剂KU60019 辐射旁效应 HEPG2细胞 

分 类 号：R735.7[医药卫生—肿瘤]