DNA糖基化酶1对幽门螺杆菌致胃上皮细胞DNA损伤的保护作用

doi:10.3969/j.issn.1004-616x.2018.02.003

癌变·畸变·突变 ›› 2018, Vol. 30 ›› Issue (2): 92-97,102.doi: 10.3969/j.issn.1004-616x.2018.02.003

DNA糖基化酶1对幽门螺杆菌致胃上皮细胞DNA损伤的保护作用

王媛¹, 邓小飞²

1. 安顺市人民医院消化内科, 贵州安顺 561000;
2. 第三军医大学新桥医院急诊科, 重庆 400037

收稿日期:2017-08-09 修回日期:2017-10-18 出版日期:2018-03-30 发布日期:2018-03-30
通讯作者: 邓小飞,E-mail:15808039698@163.com E-mail:15808039698@163.com
作者简介:王媛,E-mail:83311149@qq.com

Effect of DNA glycosylase OGG1 against Helicobacter pylori-induced DNA damage in gastric epithelial cells

WANG Yuan¹, DENG Xiaofei²

1. Department of Gastroenterology, Anshun Municipal People's Hospital, Anshun 561000, Guizhou;
2. Department of Emergency, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China

Received:2017-08-09 Revised:2017-10-18 Online:2018-03-30 Published:2018-03-30

摘要/Abstract

摘要： 目的：探讨氧化性DNA损伤修复关键酶DNA糖基化酶1（OGG1）在幽门螺杆菌（HP）感染致胃上皮细胞（GES-1） DNA损伤中的作用。方法：实验分为对照组、HP感染组、OGG1干扰组、HP感染组+OGG1干扰组。采用RNA干扰技术沉默GES-1细胞OGG1表达，24 h后进行HP感染，于感染后24 h和48 h分别采用CCK-8试验和LDH释放试验检测细胞活力和细胞坏死情况；感染48 h后分别采用彗星试验和γH2AX焦点形成试验检测细胞DNA损伤；采用TUNEL试验和PARP表达检测细胞凋亡情况。结果：与对照组相比，HP感染组和单纯OGG1干扰组的细胞活力、LDH水平、DNA损伤情况、γH2AX焦点形成、TUNEL阳性细胞数目及活性PARP表达水平均无显著变化（P均 > 0.05）。而OGG1干扰+HP感染则引起胃上皮细胞活力下降、LDH释放增加、DNA损伤加重、γH2AX焦点形成增多、TUNEL阳性细胞增多及活性PARP表达升高，与对照组及HP感染组相比，差异均有统计学意义（P < 0.05或0.01）。结论：OGG1在HP感染致胃上皮细胞氧化性DNA损伤中发挥了保护作用，为防护HP感染相关的胃部疾病提供了新的策略和方向。

关键词: DNA糖基化酶1, 幽门螺杆菌, DNA损伤, γH2AX焦点, 氧化应激

Abstract: OBJECTIVE: DNA glycosylase 1 (OGG1) is a critical factor involved in oxidative DNA damage repair,the aim of this study was to investigate the role of OGG1 in Helicobacter pylori (HP)-induced DNA damage in gastric epithelial cells (GES-1). METHODS: Cultured cells were divided into four treatment groups:control,HP infection,OGG1 siRNA,HP infection plus OGG1 siRNA. Human gastric epithelial cells (GES-1) were infected with HP at 24 h after OGG1 inhibition through the use of RNA interference. 24 h and 48 h after HP infection,cytotoxicity was evaluated based on viability using the CCK-8 assay and lactate dehydrogenase (LDH) release. DNA damage was measured using the phosphorylated H2AX (γH2AX) and the comet assays. Apoptosis was determined by TUNEL assay and western blot against poly-ADP-ribose polymerase (PARP). RESULTS: Compared to the control,no significant differences were found in cell viability,LDH release,DNA damage,γH2AX foci formation,TUNEL-positive cells and the expression levels of cleaved PARP in HP infection and OGG1 siRNA groups. However,compared to the control and the HP infection groups,HP infected cells with OGG1 deficiency showed reduction in cell viability and increased LDH release. Moreover,HP infection in OGG1 deficient cells induced DNA damage and γH2AX foci formation,elevated TUNEL-positive cells and expression of cleaved PARP. After statistical analysis,there were significant differences in the above indexes (P < 0.05 or 0.01). CONCLUSION: These findings indicate that OGG1 played a protective role in HP-induced DNA damage in gastric epithelial cells and the information may provide a new strategy for preventing HP infection-related gastric diseases.

Key words: DNA glycosylase 1, Helicobacter pylori, DNA damage, γH2AX focus, oxidative stress

中图分类号:

R730.231

王媛, 邓小飞. DNA糖基化酶1对幽门螺杆菌致胃上皮细胞DNA损伤的保护作用[J]. 癌变·畸变·突变, 2018, 30(2): 92-97,102.

WANG Yuan, DENG Xiaofei. Effect of DNA glycosylase OGG1 against Helicobacter pylori-induced DNA damage in gastric epithelial cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(2): 92-97,102.

参考文献

[1] SHIMIZU T,CHIBA T,MARUSAWA H. Helicobacter pylori-mediated genetic instability and gastric carcinogenesis[M]//Molecular Pathogenesie and Signal Transduction by Helicobacter pylori. Springer Interntatienal Publishing,2017:305-323.
[2] KALISPERATI P,SPANOU E,PATERAS I S,et al. Inflammation,DNA damage,helicobacter pylori and gastric tumorigenesis[J]. Front Genet,2017,8:20.
[3] GOBERT A P,WILSON K T. Polyamine-and NADPH-dependent generation of ROS during Helicobacter pylori infection:A blessing in disguise[J]. Free Radic Biol Med,2017,105(1):16-27.
[4] KOHNO Y,YAMAMOTO H,HIRAHASHI M,et al. Reduced MUTYH,MTH1,and OGG1 expression and TP53 mutation in diffuse-type adenocarcinoma of gastric cardia[J]. Hum Pathol,2016,52(2):145-152.
[5] RAZA Y,KHAN A,FAROOQUI A,et al. Oxidative DNA damage as a potential early biomarker of Helicobacter pylori associated carcinogenesis[J]. Pathol Oncol Res,2014,20(4):839-846.
[6] OHNISHI S,MA N,THANAN R,et al. DNA damage in inflammation-related carcinogenesis and cancer stem cells[J]. Oxid Med Cell Longev,2013,2013:387014.
[7] DEN HARTOG G,CHATTOPADHYAY R,ABLACK A,et al. Regulation of Rac1 and reactive oxygen species production in response to infection of gastrointestinal epithelia[J]. PLoS Pathog,2016,12(1):e1005382.
[8] JANG S H,LIM J W,MORIO T,et al. Lycopene inhibits Helicobacter pylori-induced ATM/ATR-dependent DNA damage response in gastric epithelial AGS cells[J]. Free Radic Biol Med,2012,52(3):607-615.
[9] AYALA-PENA S. Role of oxidative DNA damage in mitochondrial dysfunction and Huntington's disease pathogenesis[J]. Free Radic Biol Med,2013,62:102-110.
[10] IDELCHIK M,BEGLEY U,BEGLEY T J,et al. Mitochondrial ROS control of cancer[C]//Seminars in Cancer Biology. Academic Press,2017.
[11] RUCHKO M V,GORODNYA O M,ZULETA A,et al. The DNA glycosylase Ogg1 defends against oxidant-induced mtDNA damage and apoptosis in pulmonary artery endothelial cells[J]. Free Radic Biol Med,2011,50(9):1107-1113.
[12] DAS S,NATH S,BHOWMIK A,et al. Association between OGG1 Ser326Cys polymorphism and risk of upper aero-digestive tract and gastrointestinal cancers:a meta-analysis[J]. Springerplus,2016,5:227.
[13] SANTOS J C,FUNCK A,SILVA-FERNANDES I J,et al. Effect of APE1 T2197G (Asp148Glu) polymorphism on APE1,XRCC1,PARP1 and OGG1 expression in patients with colorectal cancer[J]. Int J Mol Sci,2014,15(10):17333-17343.
[14] RAMANIUK V P,NIKITCHENKO N V,SAVINA N V,et al. Polymorphism of DNA repair genes OGG1,XRCC1,XPD and ERCC6 in bladder cancer in Belarus[J]. Biomarkers,2014,19(6):509-516.
[15] YAN Y,CHEN X,LI T,et al. Association of OGG1 Ser326Cys polymorphism and pancreatic cancer susceptibility:evidence from a meta-analysis[J]. Tumor Biol,2014,35(3):2397-2402.
[16] SAKUMI K,TOMINAGA Y,FURUICHI M,et al. Ogg1 knockout-associated lung tumorigenesis and its suppression by Mth1 gene disruption[J]. Cancer Res,2003,63(5):902-905.
[17] YUZEFOVYCH L V,KAHN A G,SCHULER M A,et al. Mitochondrial DNA repair through OGG1 activity attenuates breast cancer progression and metastasis[J]. Cancer Res,2016,76(1):30-34.

DNA糖基化酶1对幽门螺杆菌致胃上皮细胞DNA损伤的保护作用

Effect of DNA glycosylase OGG1 against Helicobacter pylori-induced DNA damage in gastric epithelial cells

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