沉默H2AX食管癌ECA109中MDC1和53BP1斑点的变化  

The impact to MDC1 and 53BP1 after silence H2AX in esophageal carcinomas ECA109

作　　者：史鸿云[1,2] 祝淑钗[1] 刘志坤[1] 苏景伟[1] 

机构地区：[1]河北医科大学第四医院放疗科,河北石家庄050017 [2]河北大学附属医院放疗科,河北保定071000

出　　处：《基础医学与临床》2013年第7期808-813,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30870743)

摘　　要：目的探讨沉默H2AX后在食管高分化鳞状细胞癌ECA109细胞中应答电离辐射时对MDC1和53BP1的影响。方法构建沉默H2AX的慢病毒载体,将慢病毒转染食管癌ECA109细胞并检测转染后的沉默效用;免疫荧光检测r-H2AX、MDC1和53BP1核内斑点的情况以及用Western blot检测这几种蛋白的表达。结果 1)成功构建了沉默H2AX的ECA109细胞。2)电离辐射可引起r-H2AX表达量的增加,不引起MDC1和53BP1表达的增加,同时电离辐射诱导产生的r-H2AX、MDC1和53BP1核内斑点变化的规律一致。3)沉默H2AX后的ECA109细胞核内r-H2AX、MDC1和53BP1斑点的数量明显减少,尽管MDC1和53BP1蛋白表达无变化。结论在ECA109细胞中H2AX是电离辐射后比较早的反应蛋白,可以调节下游MDC1和53BP1斑点的位置。Objective To discover the impact to response of MDC1 and 53BP1, after silence H2AX to ionizing ra- diation of esophageal carcinoma ECA109. Methods Construction of lentiviral vector silencing H2AX, then trans- fecting lentivirus to cell line ECA109 and to detect the silence effect after transfection. Immunofluorescence detec- tion the nuclear focus of r-H2AX,MDC1 and 53BP1 in ECA109 with ionizing radiation before and after transfec- tion. As well as detection the expression of three proteins by western blot. Results 1 ） Successful silence of H2AX in ECA109 cells. 2）Ionizing radiation increased r-H2AX expression but not MDC1 and 53BP1. Ionizing radiation- induced nuclear focus of r-H2AX, MDCI and 53BP1 are similar. 3）The nucleus spots of r-H2AX, MDC1 and 53BP1 are significantly reduced in ECA109 after silence H2AX. Protein expression did not change. Conclusions H2AX is one of early ionizing radiation-reaction protein in ECA109 and function as the damage response at up- stream and can adjust the position of MDCI and 53BP1.

关 键 词：食管癌 r-H2AX MDC1 53BP1 核内斑点 

分 类 号：R735.1[医药卫生—肿瘤]