An investigation of trichloroethylene-induced effects on histone methylation in L-02 hepatic cells
10.3760/cma.j.issn.0253-9624.2017.04.013
- VernacularTitle: 三氯乙烯对人正常肝细胞组蛋白质甲基化修饰影响的研究
- Author:
Rongxia DENG
1
;
Xiaohu REN
;
Jiawen RUAN
;
Jian ZHENG
;
Jiacheng ZHONG
;
Weixue LU
;
Xiaoqing ZOU
;
Jianjun LIU
Author Information
1. The Department of Chemistry, Xiangtan University, Xiangtan 411105, China
- Publication Type:Journal Article
- Keywords:
Trichloroethylene;
Histones;
Normal human liver cells;
DNA damage response
- From:
Chinese Journal of Preventive Medicine
2017;51(4):347-352
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To further explore TCE-induced hepatotoxicity and its mechanisms by identification of trichloroethylene (TCE) induced abnormal histone methylation in human liver cells.
Methods:L-02 cells were treated with 0 and 8 mmol/L TCE for 24 h. Histones were extracted by acid. Liquid chromatography electrospray ionization tandem mass spectrometry (ESI-LC-MS/MS) were used to identify and quantify TCE related histone methylations. TCE induced abnormal methylation of H3K79 me2 and H3K79 me3 were validated by Western blot analysis. The further analysis of the function of histone abnormal methylation modifications were done by single cell gel electrophoresis (SCGE) and Western blot analysis of p53 and ɤH2AX.
Results:After treatment with TCE for 24 h in L-02 cells, the 36 TCE related histone methylation sites in 28 peptide segments were identified by MS. After treatment with TCE in concentrations of 0 and 8.0 mmol/L in L-02 cells for 24 h, the relative expression level of histone H3K79 me3 were 1.00±0.06, 0.70±0.09 (t=15.01, P=0.015); the relative expression level of histone H3K79 me2 were 1.00±0.05, 0.74±0.07 (t=16.69, P=0.018); the Olive Tail Moment about DNA damage were 1.46±0.28, 3.12± 0.68 (t=15.22, P=0.018); the relative expression levels of p53 were 1.00±0.04, 1.24±0.04 (t=18.71, P= 0.012); and the relative expression levels of ɤH2AX were 1.00 ± 0.03, 1.56 ± 0.11 (t=8.32, P=0 045).
Conclusion:TCE can induce changes in the relative expression level of H3K79 me2 and H3K79 me3 in L-02 cell, and induce DNA damage, suggesting that TCE may induce changes in the relative expression level of H3K79 me2 and H3K79 me3 by DNA damage.
