[1]张志敏,杨镇洲,何昊,等.APE1乙酰化调节电离辐射诱导的HeLa细胞自噬[J].第三军医大学学报,2016,38(04):380-384.
 Zhang Zhimin,Yang Zhenzhou,He Hao,et al.APE1 acetylation regulates autophagy induced by ionizing radiation in HeLa cells[J].J Third Mil Med Univ,2016,38(04):380-384.
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APE1乙酰化调节电离辐射诱导的HeLa细胞自噬(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第04期
页码:
380-384
栏目:
基础医学
出版日期:
2016-02-29

文章信息/Info

Title:
APE1 acetylation regulates autophagy induced by ionizing radiation in HeLa cells
作者:
张志敏杨镇洲何昊蓝保华李钱吴艳王帅李娟
第三军医大学大坪医院野战外科研究所肿瘤中心
Author(s):
Zhang Zhimin Yang Zhenzhou He Hao Lan Baohua Li Qian Wu Yan Wang Shuai Li Juan

Cancer Center, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042, China

关键词:
电离辐射APE1乙酰化自噬宫颈癌
Keywords:
ionizing radiation APE1 acetylation autophagy cervical cancer
分类号:
R341;R730.23;R737.33
文献标志码:
A
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HeLa细胞自噬的调节作用。      方法      给予Elekta precise直线加速器处理HeLa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      HeLa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)。HeLa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HeLa细胞自噬。

Abstract:

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/K7R cells and APE1WT cells underwent radiation treatment with Elekta precise linear accelerator, and APE1 acetylation and interaction of Beclin1 and Bcl2 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). LC3Ⅱ 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 LC3Ⅱ expression was up-regulated (P<0.05). Compared with APE1WT cells, the interaction of Beclin1 and Bcl2 was enhanced in APE1K6R/K7R cells, LC3Ⅱ/LC3Ⅰ ratio 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 Beclin1/Bcl2 pathway in HeLa cells.

参考文献/References:

[1]Raffoul J J, Banerjee S, Singh-Gupta V, et al. Down-regulation of apurinic/apyrimidinic endonuclease 1/redox factor-1 expression by soy isoflavones enhances prostate cancer radiotherapy in vitro and in vivo[J]. Cancer Res, 2007, 67(5): 2141-2149. DOI:10.1158/0008-5472.CAN-06-2147
[2]Robertson K A, Bullock H A, Xu Y, et al. Altered expression of Ape1/ref-1 in germ cell tumors and overexpression in NT2 cells confers resistance to bleomycin and radiation[J]. Cancer Res, 2001, 61(5): 2220-2225.
[3]Poletto M, Di-Loreto C, Marasco D, et al. Acetylation on critical lysine residues of Apurinic/apyrimidinic endonuclease 1 (APE1) in triple negative breast cancers[J]. Biochem Biophys Res Commun, 2012, 424(1): 34-39. DOI:10.1016/j.bbrc.2012.06.039
[4]Wang Y T, Tzeng D W, Wang C Y, et al. APE1/Ref-1 prevents oxidative inactivation of ERK for G1-to-S progression following lead acetate exposure[J]. Toxicology, 2013, 305: 120-129. DOI:10.1016/j.tox.2013.01.010
[5]Yamamori T, DeRicco J, Naqvi A, et al. SIRT1 deacetylates APE1 and regulates cellular base excision repair[J]. Nucleic Acids Res, 2010, 38(3): 832-845. DOI:10.1093/nar/gkp1039
[6]Bhakat K K, Izumi T, Yang S H, et al. Role of acetylated human AP-endonuclease (APE1/Ref-1) in regulation of the parathyroid hormone gene[J]. EMBO J, 2003, 22(23): 6299-6309. DOI:10.1093/emboj/cdg595
[7]Munoz-Gamez J A, Rodriguez-Vargas J M, Quiles-Perez R, et al. PARP-1 is involved in autophagy induced by DNA damage[J]. Autophagy, 2009, 5(1): 61-74.
[8]Lirussi L, Antoniali G, Vascotto C, et al. Nucleolar accumulation of APE1 depends on charged lysine residues that undergo acetylation upon genotoxic stress and modulate its BER activity in cells[J]. Mol Biology Cell, 2012, 23(20): 4079-4096. DOI:10.1091/mbc.E12-04-0299
[9]Liang N, Jia L, Liu Y, et al. ATM pathway is essential for ionizing radiation-induced autophagy[J]. Cell Signal, 2013, 25(12): 2530-2539. DOI:10.1016/j.cellsig.2013.08.010
[10]Pan Y Z, Wang X, Bai H, et al. Autophagy in drug resistance of the multiple myeloma cell line RPMI8226 to doxorubicin[J]. Genet Mol Res, 2015, 14(2): 5621-5629. DOI:10.4238/2015.May.25.14
[11]Shan J L, He H T, Li M X, et al. APE1 promotes antioxidant capacity by regulating Nrf-2 function through a redox-dependent mechanism[J]. Free Radic Biol Med, 2015, 78: 11-22. DOI:10.1016/j.freeradbiomed.2014.10.007
[12]Zhao J, Gao F, Zhang Y, et al. Bcl2 inhibits abasic site repair by down-regulating APE1 endonuclease activity[J]. J Biol Chem, 2008, 283(15): 9925-9932. DOI:10.1074/jbc.M708345200

更新日期/Last Update: 2016-01-29