纳米氧化石墨烯诱导人类支气管上皮细胞DNA氧化与甲基化的实验研究

doi:10.3969/j.issn.1004-616x.2019.06.007

癌变·畸变·突变 ›› 2019, Vol. 31 ›› Issue (6): 454-458,463.doi: 10.3969/j.issn.1004-616x.2019.06.007

纳米氧化石墨烯诱导人类支气管上皮细胞DNA氧化与甲基化的实验研究

彭长凤, 谢杏, 柯跃斌

深圳市疾病预防控制中心, 广东深圳 518020

收稿日期:2019-08-01 修回日期:2019-10-23 出版日期:2019-11-30 发布日期:2019-12-04
通讯作者: 柯跃斌,E-mail:keyke@szu.edu.cn E-mail:keyke@szu.edu.cn
作者简介:彭长凤,E-mail:907538362@qq.com。
基金资助:
国家自然科学基金项目（81072323/H2607）；深圳市科委基础研究项目（JCYJ20170306161056974）；深圳市三名工程项目（SZSM20 1611068）

DNA oxidation and methylation effects in nano-graphene oxide-exposed 16HBE cells

PENG Changfeng, XIE Xing, KE Yuebin

Shenzhen Center for Diseases Control and Prevention, Shenzhen 518020, Guangdong, China

Received:2019-08-01 Revised:2019-10-23 Online:2019-11-30 Published:2019-12-04

摘要/Abstract

摘要： 目的：通过分析体外培养的人类支气管上皮细胞（16HBE）在纳米氧化石墨烯（NGO）染毒后细胞DNA中8-羟基脱氧鸟嘌呤核苷（8-OHdG）和5-甲基胞嘧啶（5-mC）比例的变化规律，探讨DNA氧化与DNA甲基化的关联性。方法：分别以1.25、2.5、5、10 μg/mL的NGO溶液染毒16HBE细胞24 h，采用高效液相色谱串联电化学检测技术分析细胞DNA中8-OHdG比例，高效毛细管电泳（HPCE）法检测细胞全基因组DNA甲基化水平。结果：在2.5、5、10 μg/mL的NGO溶液染毒后，细胞DNA中8-OHdG比例显著高于对照组（P < 0.05或P < 0.01），以10 μg/mL浓度时为最高。NGO可引起16HBE细胞甲基化程度降低，与对照组细胞相比，2.5、5和10 μg/mL NGO组分别降低37.1%、49.7%和46.3%，与对照组间的差异均有统计学意义（P < 0.05或P < 0.01）。DNA氧化水平与DNA甲基化水平间呈负相关（r=-0.69，P < 0.01）。结论：本研究提示，在体外NGO可导致人支气管上皮细胞16HBE的DNA氧化水平升高，DNA甲基化水平降低，我们推测细胞DNA氧化程度可能影响DNA甲基化过程。

关键词: 纳米氧化石墨烯, DNA氧化, 8-羟基脱氧鸟苷, DNA甲基化, 去甲基化

Abstract: OBJECTIVE: To study the induction of DNA oxidation and methylation in nano-graphene oxide (NGO)-exposed 16HBE cells,the quantities of 8-OHdG and DNA 5-mC were determined. METHODS: 16HBE cells were exposed to 1.25,2.5,5 or 10 μg/mL NGO solution for 24 h,8-OHdG and genomic DNA 5-mC were analyzed using high-performance liquid chromatography and high performance capillary electrophoresis,respectively. RESULTS: NGO induced the formation of 8-OHdG and 5-mC in different characteristics. With 2.5,5,10 μg/mL NGO,8-OHdG quantities in the treated groups were significantly higher than that in the control group (P < 0.05 or P < 0.01),and in a dose-dependent manner. However,significant reduction of DNA methylation was observed using the same doses,by 37.1%,49.7% and 46.3%,respectively,compared to that in the control group (P < 0.05 or P < 0.01). The two endpoints were significantly and negatively correlated (r=-0.69,P < 0.01). CONCLUSION: Our data indicate that NGO exposure in cells caused increased DNA oxidation but decreased DNA methylation. It is possible that cellular DNA oxidation may affect DNA methylation.

Key words: nano-graphene oxide, DNA oxidation, 8-OHdG, DNA methylation, demethylation

中图分类号:

Q355

彭长凤, 谢杏, 柯跃斌. 纳米氧化石墨烯诱导人类支气管上皮细胞DNA氧化与甲基化的实验研究[J]. 癌变·畸变·突变, 2019, 31(6): 454-458,463.

PENG Changfeng, XIE Xing, KE Yuebin. DNA oxidation and methylation effects in nano-graphene oxide-exposed 16HBE cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(6): 454-458,463.

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纳米氧化石墨烯诱导人类支气管上皮细胞DNA氧化与甲基化的实验研究

DNA oxidation and methylation effects in nano-graphene oxide-exposed 16HBE cells

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