不同表面修饰的氧化铁纳米颗粒对A549细胞的毒性及DNA损伤

doi:10.3969/j.issn.1004-616x.2018.06.001

癌变·畸变·突变 ›› 2018, Vol. 30 ›› Issue (6): 413-421.doi: 10.3969/j.issn.1004-616x.2018.06.001

• 论著 • 下一篇

不同表面修饰的氧化铁纳米颗粒对A549细胞的毒性及DNA损伤

文海若¹, 郭雅娟^1,2, 黄芝瑛², 王雪¹, 淡墨¹

1. 中国食品药品检定研究院国家药物安全评价监测中心, 药物非临床安全评价研究北京市重点实验室, 北京 100176;
2. 中山大学药学院, 广东广州 510006

收稿日期:2018-06-29 修回日期:2018-11-13 出版日期:2018-11-30 发布日期:2018-11-30
通讯作者: 王雪,E-mail:xue_wang@nifdc.org.cn;淡墨,E-mail:danmo@nifdc.org.cn E-mail:xue_wang@nifdc.org.cn;danmo@nifdc.org.cn
作者简介:文海若,E-mail:wenhairuo@nifdc.org.cn;郭雅娟,E-mail:1141023662@qq.com。
基金资助:
国家自然科学基金（81401517）

Induction of cytotoxicity and DNA damage by iron oxide nanoparticles with different surface modifications in A549 cells

WEN Hairuo¹, GUO Yajuan^1,2, HUANG Zhiying², WANG Xue¹, DAN Mo¹

1. National Center for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, Key Laboratory of Beijing for Nonclinical Safety Evaluation Research of Drugs, Beijing 100176;
2. School of Phamaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China

Received:2018-06-29 Revised:2018-11-13 Online:2018-11-30 Published:2018-11-30

摘要/Abstract

摘要： 目的：比较不同表面修饰的氧化铁纳米颗粒（IONPs）对A549细胞的细胞毒性、染色体和DNA损伤及其作用机制的差异。方法：比较相同粒径范围（约5 nm）的胺基表面修饰的纳米氧化铁颗粒（Amine-IONP）和聚乙二醇表面修饰的纳米氧化铁颗粒（PEG-IONP）对A549细胞存活率和细胞周期的影响；使用高内涵法检测细胞经IONPs处理后的胞内活性氧簇（ROS）含量、线粒体膜电位（MMP）和内质网（ER）状态的变化；采用体外胞质分裂法微核试验和彗星电泳评价IONPs对染色体和DNA完整性的影响。结果：两种纳米氧化铁颗粒处理48 h对A549细胞生长抑制率均小于20%。相同浓度条件下，PEG-IONP主要表现为对A549细胞G0/G1期阻滞，自20 μg/mL起即明显减少S期细胞比率（P < 0.01），320 μg/mL PEG-IONP处理24 h后可诱导p21与p53表达水平显著升高（P < 0.05）。给药48 h时，Amine-IONP作用后细胞ROS、MMP及ER水平显著性改变的起始浓度分别为20、20和80 μg/mL，而PEG-IONP作用后产生显著性改变的起始浓度分别为40、40和160 μg/mL。此外，与PEG-IONP比较，Amine-IONP可在较低浓度条件下诱导微核和彗星拖尾形成（Amine-IONP的起始浓度为20和80 μg/mL；而PEG-IONP则为40和160 μg/mL）。经氧自由基清除剂乙酰半胱氨酸和叔丁基对羟基茴香醚预处理后，两者导致的胞内ROS含量和尾部DNA百分率均明显降低（P < 0.05）。结论：带正电荷的Amine-IONP更易于诱导A549细胞氧化应激及与之有关的DNA损伤；相比之下，PEG-IONP的细胞毒性和遗传毒性较弱，但除氧化损伤外也可通过抑制细胞周期干扰细胞增殖，作为肿瘤诊断试剂具有一定优势。

关键词: 氧化铁纳米颗粒, A549细胞, 微核试验, 彗星试验, 氧化应激, 细胞增殖

Abstract: OBJECTIVE: To compare the induction of cytotoxicity,chromosome aberrations and DNA damage among iron oxide nanoparticles (IONPs) with different surface-modifications in A549 cells. METHODS: The effects of amine-modified (Amine-IONP) and polyethylene glycol-modified iron oxide nanoparticles (PEG-IONP) of the same particle size range (5 nm) on cell viability and cell cycle were compared in A549 cells;the content of intracellular reactive oxygen species (ROS),mitochondrial membrane potential (MMP) and endoplasmi reticulum stress (ERS) in cells treated with IONPs were analyzed using high content screening method;the in vitro cytokinesis micronucleus test and Comet assay were performed to evaluate their effects on chromosome and DNA integrity. RESULTS: The growth inhibitory rates of both IONPs to A549 cells were less than 20% after 48 h. Under the same concentration,PEG-IONP exhibited a more significant effect on the G0/G1 cell cycle arrest (P < 0.05):the initial concentration to significantly reduce the cell rate of S phase was 20 μg/mL,and the expression of p21 and p53 was up-regulated by a treatment of 320 μg/mL PEG-IONP for 24 h (P < 0.05). After a 48 h treatment,the initial concentrations to show significant effects of Amine-IONP on ROS,MMP and ER were 20,20 and 80 μg/mL,while for PEG-IONP they were 40, 40 and 160 μg/mL respectively. In addition,compared to PEG-IONP,Amine-IONP was able to induce the formation of micronuclei and Comet tail (the initial concentrations for Amine-IONP were 20 and 80 μg/mL;while PEG-IONP were 40 and 160 μg/mL respectively). After the pre-treatment with oxygen free radical scavengers N-acetylcysteine and butylated hydroxyanisole,both IONPs induced ROS and Comet DNA Tail were significantly inhibited (P < 0.05). CONCLUSION: Positively charged Amine-IONP was more effective in inducing oxidative stress and DNA damage in A549 cells;whereas,PEG-IONP with relatively weaker cytotoxicity and genotoxicity interfered with cell proliferation,and showed advantages in the development of tumor diagnostic reagents.

Key words: iron oxide nanoparticles, A549 cells, micronucleus assay, Comet assay, oxidative stress, cell proliferation

中图分类号:

R917

文海若, 郭雅娟, 黄芝瑛, 王雪, 淡墨. 不同表面修饰的氧化铁纳米颗粒对A549细胞的毒性及DNA损伤[J]. 癌变·畸变·突变, 2018, 30(6): 413-421.

WEN Hairuo, GUO Yajuan, HUANG Zhiying, WANG Xue, DAN Mo. Induction of cytotoxicity and DNA damage by iron oxide nanoparticles with different surface modifications in A549 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(6): 413-421.

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不同表面修饰的氧化铁纳米颗粒对A549细胞的毒性及DNA损伤

Induction of cytotoxicity and DNA damage by iron oxide nanoparticles with different surface modifications in A549 cells

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