摘要
目的探讨小分子干扰RNA(siRNA)沉默BAZ1A基因对肺腺癌A549细胞的放疗增敏作用。方法将A549细胞随机分为3组:空白对照组(Ctrl)、siRNA对照组(siNC)、BAZ1A siRNA转染组(siBAZ1A),Western blot检测转染后BAZ1A蛋白表达水平;平板克隆实验检测不同辐射剂量下细胞的存活分数(SF),并通过单击多靶模型拟合细胞存活曲线,计算放射增敏比(SER);MTS、划痕实验及流式细胞仪分别检测干预后A549细胞的增殖能力、迁移能力以及细胞凋亡率。结果siBAZ1A可降低BAZ1A蛋白的表达水平,增强放疗对A549细胞克隆形成能力的抑制作用,在2~10 Gy的放射剂量下,siBAZ1A组细胞的SF值均低于其余两组,差异具有统计学意义(P<0.05),siBAZ1A组及siNC组相对于Ctrl组的SER分别为1.06和2.24,siBAZ1A组照射后,细胞增殖能力和细胞迁移能力较对照组减弱,细胞凋亡率增加,差异具有统计学意义(P<0.05)。结论沉默BAZ1A基因对肺腺癌A549细胞具有放射增敏作用。
Objective To investigate the effect of siRNA silencing BAZ1A on radiosensitization of human lung adenocarcinoma A549 cells.Methods A549 cells were randomly divided into transfection reagent control(Ctrl)group,negative control siRNA(siNC)group,and siBAZ1A group.The expression of BAZ1A protein was evaluated by Western blot.The clone formation assay was applied to investigate the survival fraction(SF)of the A549 cells treated with different radiation doses(0,2,4,6,8,and 10Gy),and one-hit multi-target model was applied to analyze the radiation dose survival curve.MTS assay,scratch assay,and flow cytometry were utilized to determine the relative survival,cell migration abilities,and apoptosis,respectively.Results The expression level of BAZ1A protein in A549 cells was inhibited by BAZ1A-siRNA transfection.X-ray radiation inhibited the colony formation capacity of A549 cells,and the SF of siBAZ1A group was lower than that of the other two groups with radiation(P<0.05).Compared with those of Ctrl group,the sensitization enhancement ratio(SER)of siNC and siBAZ1A groups was 1.06 and 2.24.Moreover,with the transfection of BAZ1A-siRNA,the relative survival rate and cell migration ability were decreased after X-ray radiation.Besides,the apoptosis rate was higher in siBAZ1A group(P<0.05).Conclusion Silencing the expression of BAZ1A by siRNA can efficiently improve the sensitization of radiotherapy for A549 cells.
作者
罗真真
张钰洋
高诚宜
张天
陈依玲
郑文芳
李俊俊
朱青
韩苏夏
LUO Zhen-zhen;ZHANG Yu-yang;GAO Cheng-yi;ZHANG Tian;CHEN Yi-ling;ZHENG Wen-fang;LI Jun-jun;ZHU Qing;HAN Su-xia(Department of Oncology Radiotherapy,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061;Department of Abdominal Oncology,West China Hospital of Sichuan University,Chengdu 610041,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2020年第1期13-17,共5页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
陕西省创新人才推进计划-重点科技创新团队(No.2018TD-002)
国家自然科学基金面上项目(No.81472795,81272488,81874223)
省级大学生创新性实验项目(No.SJ201810698142)~~
