ZEB1通过上调ATM表达调控胃癌细胞AGS的放射敏感性  

ZEB1regulates the radiosensitivity of gastric cancer cell AGS by up-regulating ATM expression

作　　者：崔勇霞[1] 吴广银[2] 罗执芬[1] 白冰[1] 陈曦 王朝杰[1] Cui Yongxia;Wu Guangyin;Luo Zhifen;Bai Bing;Chen Xi;Wang Zhaojie(Department of Oncology,Henan Provincial People's Hospital,Zhengzhou 450003,China;Department of Radiotherapy,Henan Provincial People's Hospital,Zhengzhou 450003,China)

机构地区：[1]河南省人民医院肿瘤科,郑州450003 [2]河南省人民医院放疗科,郑州450003

出　　处：《中华放射医学与防护杂志》2018年第12期894-898,共5页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金(U1204818).

摘　　要：目的 探究锌指结构E-box结合蛋白1(ZEB1)对胃癌细胞AGS放射敏感性的影响并探究其可能的作用机制。方法 通过不同剂量(0、2、4、6、8 Gy)X射线照射AGS细胞,蛋白质印迹法(Western blot)实验观测细胞中ZEB1的表达量;取对数生长期AGS细胞,将过表达ZEB1、ZEB1干扰表达质粒以及相应的对照质粒(pcDNA3.1)和阴性对照干扰质粒转染至AGS细胞中,分别记为过表达ZEB1组、沉默ZEB1组、对照组和阴性对照组,细胞克隆实验检测过表达和沉默ZEB1对AGS细胞存活率的影响;流式细胞术实验检测细胞的凋亡率;Western blot检测细胞损伤相关组蛋白H2A(H2AX)、磷酸化H2AX(γ-H2AX)以及毛细血管扩张突变基因(ATM)表达量。结果 ZEB1在AGS细胞中的表达对放射剂量具有依赖性(F=58.57,P<0.05);过表达ZEB1能够提高AGS细胞的存活,抑制γ-H2AX表达(t=12.18,P<0.05),并阻碍细胞的凋亡(t=7.27,P<0.05),并随时间上调ATM表达量(F=165.70,P<0.05);沉默ZEB1降低AGS细胞的存活,增加γ-H2AX表达(t=12.88,P<0.05)和细胞的凋亡(t=8.36,P<0.05),随时间下调ATM表达量(F=44.80,P<0.05)。结论 ZEB1通过上调ATM表达调控胃癌AGS细胞放射敏感性。Objective To investigate the effect of Zinc finger E-box binding homeobox protein 1 (ZEB1) on the radiosensitivity of gastric cancer cells AGS and its possible mechanism.Methods AGS cells were irradiated by X-rays at different doses (0, 2, 4, 6, and 8 Gy). Western blot was used to observe the expression of ZEB1 in cells. AGS cells, in logarithmic growth phase, were transfected with of ZEB1 gene or its interference plasmids, the corresponding control plasmids (pcDNA3.1) and negative control interference plasmids. They were classified as overexpression ZEB1 group, silencing ZEB1 group, control group and negative control group, respectively. The effect of overexpression and silencing ZEB1 on the survival of AGS cells after irradiation were analyzed by colony formation assay. The cell apoptosis rate was analyzed by flow cytometry. The expressions of histone H2A (H2AX), phosphorylated H2AX (γ-H2AX) and telangiectasia mutated gene (ATM) were detected by Western blot.Results The expression of ZEB1 in AGS cells was dependent on radiation dose (F=58.57, P<0.05). Overexpression of ZEB1 increased AGS cells viability, inhibited γ-H2AX expression (t=12.18, P<0.05), blocked cell apoptosis (t=7.27, P<0.05) and up-regulated ATM expression in time-dependent manner after irradaition (F=165.70, P<0.05). Silencing ZEB1 reduced AGS cells viability, increased γ-H2AX expression (t=12.88, P<0.05) and cell apoptosis (t=8.36, P<0.05), and down-regulated of ATM expression(F=44.80, P<0.05).Conclusions ZEB1 regulates the radiosensitivity of gastric cancer AGS cells by up-regulating ATM expression.

关 键 词：胃癌 放射敏感性 ZEB1 ATM 

分 类 号：R735.2[医药卫生—肿瘤]