组蛋白修饰改变在亚砷酸钠毒性效应中的作用研究

doi:10.3969/j.issn.1004-616x.2014.02.002

癌变·畸变·突变

组蛋白修饰改变在亚砷酸钠毒性效应中的作用研究

牛林梅，章征保，曾晓雯，朱小年，陈雯，李道传

中山大学公共卫生学院预防医学系，广东广州 510080

收稿日期:2014-01-06 修回日期:2014-02-21 出版日期:2014-03-30 发布日期:2014-03-30
通讯作者: 李道传，E-mail：ldc1983king@163.com
作者简介:牛林梅(1986- )，女，河南省安阳市人，硕士，研究方向：分子毒理学。E-mail：niulinmei@126.com
基金资助:
科技部973项目(2010CB912803)，广东省自然科学基金(S201204 0007713)，高等学校博士学科点专项科研基金(201201711200 67)，中山大学青年培育项目(12ykpy11)

Effect of histone modification in the toxic effects of sodium arsenite

NIU Lin-mei，ZHANG Zheng-bao，ZENG Xiao-wen，ZHU Xiao-nian，CHEN Wen，LI Diao-chuan

Faculty of Preventive Medicine, School of Public Health, Sun-Yat Sen University, Guangzhou 510080, Guangdong, China

Received:2014-01-06 Revised:2014-02-21 Online:2014-03-30 Published:2014-03-30
Contact: 李道传，E-mail：ldc1983king@163.com

摘要/Abstract

摘要：

目的：探讨组蛋白H3修饰在亚砷酸钠暴露引起的毒性效应中的作用及其调控机制。方法：在HBE细胞中构建组蛋白H3赖氨酸修饰位点突变的细胞株，用亚砷酸钠作用于细胞，四甲基噻唑蓝(MTT)法检测其细胞毒性；用微核实验检测组蛋白H3K4突变后对亚砷酸钠诱导DNA损伤的影响；用蛋白印迹检测亚砷酸钠对H3K4甲基化蛋白表达的影响并用qRT-PCR筛查了其上游修饰酶mRNA水平的表达。结果：组蛋白修饰缺陷细胞株中，H3K4修饰缺陷后的HBE细胞在1 μmol/L亚砷酸钠染毒作用下细胞活力降低60%左右(P<0.01)，且可致HBE细胞双核微核率升高2倍以上(P<0.01)；1 μmol/L亚砷酸钠处理HBE细胞可引起H3K4me2/3 蛋白水平升高(P<0.05)，同时赖氨酸特异性去甲基化修饰酶1(LSD1)的表达下降(P<0.01)。结论：亚砷酸钠染毒诱导细胞组蛋白修饰发生改变，其中H3K4甲基化修饰水平上调，可能参与砷引发的细胞毒性和遗传毒性过程，其修饰改变可能受到LSD1 的调控。

关键词: 亚砷酸钠, DNA损伤, H3K4甲基化, LSD1

Abstract:

OBJECTIVE: We attempted to investigate the function and regulation mechanism of histone H3 modifications in the toxicity induced by sodium arsenite. METHODS：We constructed histone H3 lysine modified defective cell lines by expressing the site-specific H3 mutation plasmids in HBE cells. MTT assay was used to test the cytotoxicity of these cells when exposed to sodium arsenite. Cytokinesis-block micronucleus (CBMN) assay was conducted to examine the function of H3K4 methylation on the effect of the DNA damage. Meanwhile，the HBE cells were treated with sodium arsenite and the mRNA levels of H3K4 methyltransferases and demethyltransferases and the protein levels of the H3K4 methylation were measured by qRT-PCR and Western blot assay to explore the regulation of H3K4 methylation when exposed to sodium arsenite. RESULTS：We found that the cell vitality was reduced about 60% and the rate of cytokinesis-block micronucleus increased more than 2 times when H3K4 methylation defective HBE cells were exposure to low concentrations (1 μmol/L) of sodium arsenite (P<0.01). The reduction of H3K4 methylation could increase the sensitivity of HBE cells treated with sodium arsenite. We also found that H3K4me2/3 were hypermethylated when exposed to 1 μmol/L sodium arsenite and H3K4 demethylase (lysine-specific demethylase 1，LSD1) was decreased in this process (P<0.05). CONCLUSION：The increased H3K4 methylation might be regulated by reduced LSD1，which may be involved in the cytotoxicity and genotoxity induced by sodium arsenite.

Key words: sodium arsenite, DNA damage, H3K4 metylation, lysine-specific demethylase 1

牛林梅，章征保，曾晓雯，朱小年，陈雯，李道传. 组蛋白修饰改变在亚砷酸钠毒性效应中的作用研究[J]. 癌变·畸变·突变, doi: 10.3969/j.issn.1004-616x.2014.02.002.

NIU Lin-mei，ZHANG Zheng-bao，ZENG Xiao-wen，ZHU Xiao-nian，CHEN Wen，LI Diao-chuan. Effect of histone modification in the toxic effects of sodium arsenite[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2014.02.002.

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组蛋白修饰改变在亚砷酸钠毒性效应中的作用研究

Effect of histone modification in the toxic effects of sodium arsenite

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