APE1乙酰化调节电离辐射诱导的HeLa细胞自噬  

APE1 acetylation regulates autophagy induced by ionizing radiation in HeLa cells

作　　者：张志敏[1] 杨镇洲[1] 何昊[1] 蓝保华 李钱[1] 吴艳[1] 王帅[1] 李娟[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所肿瘤中心,重庆400042

出　　处：《第三军医大学学报》2016年第4期380-384,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81272499)~~

摘　　要：目的探讨脱嘌呤/脱嘧啶核酸内切酶∕氧化还原因子(apurinic/aprimidinic endonuclease/redox factor-1,APE1/Ref-1)乙酰化对电离辐射诱导He La细胞自噬的调节作用。方法给予Elekta precise直线加速器处理He La细胞及子系细胞APE1WT和APE1K6R/K7R,应用免疫共沉淀(co-immunoprecipitation,Co-IP)的方法检测APE1乙酰化修饰及Beclin1/Bcl2的结合,流式细胞仪检测细胞自噬的水平,免疫荧光激光共聚焦检测LC3的聚集,Western blot检测APE1、LC3和p62的表达。结果 He La细胞APE1乙酰化水平随着照射剂量的增加和照射后时间的延长而逐渐升高(P<0.05)。He La细胞LC3Ⅱ的表达与照射剂量呈正相关,而与p62呈负相关(P<0.05);电离辐射促进细胞的自噬水平和LC3的聚集。电离辐射后,LC3Ⅱ上调;与对照组APE1WT相比,APE1K6R/K7R细胞Beclin1/Bcl2结合增强,LC3Ⅱ/LC3Ⅰ比值降低,p62表达上调(P<0.05)。结论 APE1乙酰化通过Beclin1/Bcl2途径调节电离辐射诱导的He La细胞自噬。Objective To explore the relationship between apurinic/aprimidinic endonuclease/redox factor-1 （APE1） acetylation and autophagy induced by ionizing radiation in HeLa cells. Methods HeLa cells, APE1K6R/KTR cells and APE1WT cells underwent radiation treatment with Elekta precise linear accelerator, and APE1 acetylation and interaction of Beclinl and Bc]2 were detected by co-immunoprecipitation （Co-IP）. Flow cytometry was used to analyze HeLa cell autophagy level, LC3 accumulation was observed by immunofluorescence assay, and Western blotting was adopted to measure the expression of APE1, LC3 and p62. Results The APE1 acetylation was increased along with the increase of the radiation dose and the time after radiation in HeLa cells, and the increase of APE1 acetylation was in a time- and dose-dependent manner （P 〈 0.05 ）. LC3 Ⅱ expression was positively correlated with the radiation dose in HeLa cells, while p62 expression was opposite （ P 〈 0. 05 ）. LC3 accumulation and autophagy increased after ionizing radiation, and the LC3 Ⅱ expression was up-regulated （P 〈0. 05）. Compared with APE1wT cells, the interaction of Beclinl and Bcl2 was enhanced in APE1K6R/KYR cells, LC3 Ⅱ/LC3 Ⅰratio was decreased, and p62 expression was up- regulated after ionizing radiation （P 〈 0. 05）. Conclusion APE1 acetylation is increased after ionizing radiation, which can promote radiation-induced autophagy via Beclinl/Bcl2 pathway in HeLa cells.

关 键 词：电离辐射 APE1乙酰化 自噬 宫颈癌 

分 类 号：R341[医药卫生—基础医学] R730.23