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

作　　者：彭长凤 谢杏[1] 柯跃斌[1] PENG Changfeng;XIE Xing;KE Yuebin(Shenzhen Center for Diseases Control and Prevention,Shenzhen 518020,Guangdong,China)

机构地区：[1]深圳市疾病预防控制中心

出　　处：《癌变．畸变．突变》2019年第6期454-458,463,共6页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：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(SZSM201611068)

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、10μg/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、10μg/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10μg/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10μg/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,D NA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。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 10μg/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 2.5,5,10μg/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.

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 

分 类 号：Q355[生物学—遗传学]