Global DNA methylation changes during palatal formation in fetal mice induced by 2,3,7,8-tetrachlrodibenzo-p-dioxin
10.3760/cma.j.issn.1009-4598.2016.05.012
- VernacularTitle:TCDD诱导的胎鼠腭发育过程中DNA甲基化变化
- Author:
Chen WANG
1
;
Xingang YUAN
;
Yuexian FU
;
Shana ZHAI
Author Information
1. 400014 重庆医科大学附属儿童医院整形外科,儿童发育疾病研究教育部重点实验室,重庆市儿科学重点实验室,重庆市(儿童发育重大疾病诊治与预防)国际科技合作基地
- Keywords:
Genome;
DNA methylation;
DNA methyltransferases;
2,3,7,8-tetrachlrodibenzo-p-dioxin;
Cleft palate
- From:
Chinese Journal of Plastic Surgery
2016;32(5):372-377
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate global DNA methylation and DNA methyhransferases participation in the mechanism of cleft palate induced by maternal exposure to 2,3,7,8-tetrachlrodibenzo-p-dioxin (TCDD)in mice.Methods 40 pregnant C57BL/6J mice were randomly divided into 2 groups:the control group(n =20) and TCDD-exposure group(n =20).On gestation day 10.5 (GD10.5),the mice in TCDD-group were orally administrated with TCDD 28 μg/kg,while the mice in the control group received equivalent corn oil.The pregnant mice were sacrificed on GD13.5,GD14.5,GD15.5,GD16.5,GD17.5,fetal palates were collected for analysis.Global DNA methylation levels were detected by MethylampTM Global DNA Methylation Quantification Ultra Kit through an ELISA-like reaction.The expression levels of DNA methyltransferases were examined by quantitative real-time PC R(q-PCR).IBM SPSS 20.0 software was applied for statistical analysis.Kolmogorov-Smirnov test was used for normal distribution check,and the distribution was normal.Independent t-test was carried out among two groups.P ＜ 0.05 was considered statistically significant.Results The global DNA methylation level in TCDD-exposure group was significantly higher than that in control group on GD13.5 (49.52％ ±4.03％ vs 33.42％ ± 6.78％,P ＜ 0.01),whilelower on GD14.5 (24.10％ ±2.29％ vs 30.12％ ±3.92％,P ＜0.05) and on GD16.5 (32.77％ ±0.98％ vs 36.45％ ± 3.27％,P ＜ 0.05).The expression level of Dnmt1 mRNA in TCDD-exposure group was higher than that in control group on GD13.5(1.28±0.11 vs 1.01 ±0.10,P＜0.05) and on GD16.5(1.04 ±0.05 vs 0.81 ±0.01,P ＜0.01).The expression level of Dnmt3a mRNA in TCDD-exposure group was higher than that in control group on GD13.5 (1.15 ±0.17 vs 0.81 ±0.02,P ＜0.05)and on GD16.5 (1.11 ± 0.06 vs 0.96 ± 0.06,P ＜ 0.05).The expression level of Dnmt3b mRNA in TCDD-exposure group was higher than that in control group on GD14.5(0.97 ±0.06 vs 0.72 ±0.06,P ＜0.01).Conclusions It is supposed that complicated mechanisms are exist to regulate global DNA methylation levels in palatal tissue of fetal mice.The significant increased DNA methylation level on GD13.5 resulting from up-expression of Dnmt1 and Dnmt3a may be one of the epigenetic mechanisms which cause palate malformation in fetal mice induced by maternal exposure to TCDD.
