三氯乙烯对B6C3F1雄性小鼠肝脏和肾脏中细胞增殖相关基因表达和DNA甲基化的影响

doi:10.3969/j.issn.1004-616x.2015.03.011

癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (3): 211-215.doi: 10.3969/j.issn.1004-616x.2015.03.011

三氯乙烯对B6C3F1雄性小鼠肝脏和肾脏中细胞增殖相关基因表达和DNA甲基化的影响

陈佳红^1,2, 张航^1,2, 赖怡^1,2, 姜岩³, 陈涛^1,2

1. 苏州大学医学部公共卫生学院, 江苏苏州 215123;
2. 江苏省老年病预防与转化医学重点实验室, 江苏苏州 215123;
3. 苏州大学医学部生物科学与基础医学学院, 江苏苏州 215123

收稿日期:2014-12-30 修回日期:2015-04-10 出版日期:2015-05-30 发布日期:2015-05-30
通讯作者: 陈涛,E-mail:tchen@suda.edu.cn E-mail:tchen@suda.edu.cn
作者简介:陈佳红,E-mail:20134247031@suda.edu.cn
基金资助:
教育部留学回国人员科研启动基金(K513900115);国家自然科学基金(81300143);江苏高校优势学科建设工程资助项目;苏州大学引进人才专项基金(Q413900512)

Cell proliferation-related mRNA expression and DNA methylation changes induced by trichloroethylene in the liver and kidney of B6C3F1 male mice

CHEN Jiahong^1,2, ZHANG Hang^1,2, LAI Yi^1,2, JIANG Yan³, CHEN Tao^1,2

1. School of Public Health, Medical College of Soochow University, Suzhou 215123;
2. Key Laboratory of Prevention and Translational Medicine Geriatrics in Jiangsu Province, Suzhou 215123;
3. School of Biology and Basic Medical Sciences, Soochow University, Suzhou 215123, Jiangsu, China

Received:2014-12-30 Revised:2015-04-10 Online:2015-05-30 Published:2015-05-30

摘要/Abstract

摘要：

目的:三氯乙烯(TCE)是环境中广泛存在的工业污染物,可引起小鼠肝癌,但对肾癌发生未见显著影响。本研究通过检测TCE对小鼠肝脏和肾脏细胞增殖和DNA甲基化调控相关基因表达以及对DNA甲基化的影响,探讨TCE引起小鼠肝癌的分子机制。方法:将6周龄B6C3F1雄性小鼠随机分成3组,每组4只,分别以0、500和1 000 mg/kg剂量的TCE连续灌胃5 d。以荧光定量PCR方法检测TCE染毒小鼠肝脏、肾脏中与细胞增殖以及DNA甲基化调控相关基因的mRNA表达水平,以结合重亚硫酸盐的限制性内切酶方法检测Cdkn1a启动子区和重复序列的DNA甲基化水平。结果:与对照组相比,TCE可引起小鼠肝脏中细胞增殖相关基因Cdkn1a、Jun和Mki67的mRNA水平显著升高(P均<0.05),且呈剂量反应关系。同时1 000 mg/kg TCE染毒小鼠肝脏中主要DNA甲基化调控基因Dnmt3a、Dnmt3b和Tet2的mRNA水平降低(P均<0.05),Uhrf1 mRNA的表达升高(P均<0.05)。TCE染毒还导致肝脏内Cdkn1a启动子区的DNA甲基化水平降低,但对肾脏中相关基因及DNA甲基化水平无显著影响。结论:TCE引起的细胞增殖相关基因表达升高及DNA甲基化异常可能在其促进小鼠肝癌发生中起重要作用。

关键词: 三氯乙烯, mRNA, DNA甲基化, 肝癌, 细胞增殖

Abstract:

OBJECTIVE: Trichloroethylene (TCE), a common industrial contaminant in the environment, could induce hepatocellular carcinoma but not kidney cancer in mice. This study aimed to understand the molecular mechanisms of TCE hepatocarcinogenicity by examining the effect of TCE on proliferation-related mRNA expression and DNA methylation in mouse liver and kidney. METHODS:B6C3F1 male mice aged six weeks were randomly divided into 3 groups, with 4 mice in each group. Mice were fed with TCE at 0, 500 and 1 000 mg/kg for 5 days. Using qPCR, we examined the mRNA expression of genes involved in cell proliferation and in the regulation of DNA methylation in TCE-exposed mouse liver and kidney. The promoter methylation status of Cdkn1a and the DNA methylation level of repetitive sequences were detected by combined bisulfite restriction analysis. RESULTS:Compared with control, TCE increased the mRNA expression of Cdkn1a, Jun and Mki67 in a dose-dependent manner in mouse liver. Moreover, in mouse treated with TCE at 1 000 mg/kg, the mRNA expression of a number of key DNA methylation regulation genes including Dnmt3a, Dnmt3b and Tet2 were down-regulated, while the mRNA expression level of Uhrf1 was up-regulated (all P<0.05). TCE also induced hypomethylation of the promoter region of Cdkn1a in mouse hepatocytes. However, TCE should no detectable effects on mRNA expression and DNA methylation in mouse kidney. CONCLUSION:Alterations in the mRNA expression of proliferation-related genes and DNA methylation changes might be crucial to TCE-induced hepatocarcinogenesis in mice.

Key words: trichloroethylene, mRNA, DNA methylation, hepatocellular carcinoma, cell proliferation

中图分类号:

R114

陈佳红, 张航, 赖怡, 姜岩, 陈涛. 三氯乙烯对B6C3F1雄性小鼠肝脏和肾脏中细胞增殖相关基因表达和DNA甲基化的影响[J]. 癌变·畸变·突变, 2015, 27(3): 211-215.

CHEN Jiahong, ZHANG Hang, LAI Yi, JIANG Yan, CHEN Tao. Cell proliferation-related mRNA expression and DNA methylation changes induced by trichloroethylene in the liver and kidney of B6C3F1 male mice[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(3): 211-215.

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三氯乙烯对B6C3F1雄性小鼠肝脏和肾脏中细胞增殖相关基因表达和DNA甲基化的影响

Cell proliferation-related mRNA expression and DNA methylation changes induced by trichloroethylene in the liver and kidney of B6C3F1 male mice

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